Wodarg Yeadon EMA Petition Pfizer Trial FINAL 01DEC2020 en Unsigned With Exhibits

PETITIONER: December 1, 2020
Dr. med. Wolfgang Wodarg
Germany
CO-PETITIONER:
Dr. Michael Yeadon
England, CT3 1RT
TO:
European Medicines Agency
Committee for human medicinal products (CHMP)
COVID-19 EMA pandemic Task Force (COVID-ETF)
Domenico Scarlattilaan 6
1083 HS Amsterdam
The Netherlands
info@ema.europa.eu
press@ema.europa.eu
!! URGENT !!
PETITION/MOTION FOR
ADMINISTRATIVE/REGULATORY ACTION REGARDING
CONFIRMATION OF EFFICACY END POINTS AND USE OF DATA IN
CONNECTION WITH THE FOLLOWING CLINICAL TRIAL(S):
PHASE III - EUDRACT NUMBER: 2020-002641-42
SPONSOR PROTOCOL NUMBER: C4591001
SPONSOR:
BIONTECH SE (SOCIETAS EUROPAEA), AN DER GOLDGRUBE 12, 55131 MAINZ,
GERMANY
AND ANY OTHER ONGOING CLINICAL TRIALS OF VACCINE CANDIDATES

DESIGNED TO STOP TRANSMISSION OF THE VIRUS FROM THE VACCINE
RECIPIENT TO OTHERS AND/OR TO PREVENT COVID-19 OR MITIGATE
SYMPTOMS OF COVID-19 FOR WHICH PCR RESULTS ARE THE PRIMARY
EVIDENCE OF INFECTION
WITH SARS-COV-2
ADMINISTRATIVE/REGULATORY STAY OF ACTION
This petition for a stay of action is submitted by the undersigned (“Petitioner” or “Lead
Petitioner”) to request the EMA a) stay the Phase III clinical trial(s) of BNT162b (EudraCT
Number 2020-002641-42) in the EU (current protocol country: Germany) until study design is
amended to conform with the requests in the “Action Requested” section (B.) below; and b) stay
all other clinical trials of vaccine candidates designed to stop transmission of the virus from the
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vaccine recipient to others and/or prevent or mitigate symptoms of COVID-19 for which PCR
results are the primary evidence of infection.
Because of the compelling need to ensure the safety and efficacy of any COVID-19 vaccine
licensed by the EMA (and/or the German Paul-Ehrlich-Institut), and to allow Petitioner the
opportunity to seek appropriate emergency judicial relief should the EMA deny its Petition,
Petitioner respectfully requests that EMA act on the instant Petition immediately.
A. DECISIONS INVOLVED
I. Approval of trial design and/or decision to not challenge trial design for Phase III trial
of BNT162 (EudraCT Number 2020-002641-42)
II. Approval of trial design and/or decision to not challenge trial design of all other
clinical trials of vaccine candidates designed to stop transmission of the virus from the vaccine
recipient to others and/or to prevent or mitigate symptoms of COVID-19 for which PCR results
are the primary evidence of infection.
B. ACTION REQUESTED
I. Stay the Phase III trial of BNT162 in the protocol country Germany and in any other
EU protocol countries (as applicable) until study design is amended to provide that:
Before an Emergency Authorization/Conditional Approval and/or Unrestricted

Authorization is issued for the Pfizer/BioNTech vaccine, all “endpoints” or COVID-19 cases
used to determine vaccine efficacy in the Phase 3 or 2/3 trials should have their infection status
confirmed by appropriate Sanger sequencing (as described in section C. III. below), given a) the
high cycle thresholds used in some trials; and b) design flaws of certain RT-qPCR tests
identical to or modeled after what is sometimes called the “Drosten-Test”.
II. Stay the clinical trials of all vaccine candidates designed to stop transmission of the
virus from the vaccine recipient to others and/or to prevent or mitigate symptoms of COVID-19
for which PCR results are the primary evidence of infection until study design is amended to
provide that:
Before an Emergency Authorization/Conditional Approval and/or Unrestricted

Authorization is issued for vaccine designed to stop transmission of the virus from the vaccine
recipient to others and/or to prevent or mitigate symptoms of COVID-19, all “endpoints” or
COVID-19 cases used to determine vaccine efficacy should have their infection status
confirmed by appropriate Sanger sequencing (as described in section B. III. below), given a) the
high cycle thresholds used in some trials; and b) design flaws of certain RT-qPCR tests
identical to or modeled after what is sometimes called the “Drosten-Test”.
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III. High cycle thresholds, or Ct values, in RT-qPCR test results have been widely
acknowledged to lead to false positives. Also, a group of scientists and researchers have
recently called for a retraction of the paper that describes the so called “Drosten-Test”
(sometimes also being referred to as the “Corman-Drosten protocol” - a specific RT-qPCR
assay described by Corman,Victor M., Drosten, Christian and others in “Detection of 2019
novel coronavirus (2019-nCoV) by real-time RT-PCR.” Euro Surveillance
2020;25(3):pii=2000045. https://doi.org/10.2807/1560-7917.ES.2020.25.3.2000045).
All RT-qPCR-positive test results used to categorize patient as “COVID-19 cases” in the trials
and used to qualify the trial’s endpoints should be verified by Sanger sequencing to confirm that
the tested samples in fact contain a unique SARS-CoV-2 genomic RNA. Congruent with FDA
and EMA requirements for a confirmed diagnosis of human papillomavirus (HPV) using PCR,
the sequencing electropherogram must show a minimum of 100 contiguous bases matching the
reference sequence with an Expected Value (E Value) <10-30 for the specific SARS-CoV-2
gene sequence based on a BLAST search of the GenBank database (aka NCBI Nucleotide
database).
C. STATEMENT OF GROUNDS
I. As detailed herein, (i) without the requested stay, the Petitioner and many EU
residents/citizens will suffer irreparable harm, (ii) the request is not frivolous and is being
pursued in good faith, (iii) the request demonstrates sound public policy, and (iv) the public
interest favors granting a stay.
II. Petitioner deems the current study designs for the Phase II/III trials of BNT162b (“the
Pfizer/BioNTech trial”) to be inadequate to accurately assess efficacy. Petitioner also deems the
designs of clinical trials of vaccine candidates designed to stop transmission of the virus from the
vaccine recipient to others and/or to prevent or mitigate symptoms of COVID-19 for which PCR
results are the primary evidence of infection to be inadequate to accurately assess efficacy.
III. Petitioner and the public will suffer irreparable harm if the actions requested herein are
not granted, because once the EMA (and other appropriate bodies in the various EU member
states) approves the COVID-19 vaccines in question, both governments of EU member states
and employers in the EU are most likely going to recommend them for widespread use. If the
assignment of cases and non-cases during the course of the trials is not accurate, the vaccines
will not have been properly tested. If the vaccines are not properly tested, important public
policy decisions regarding its use will be based on misleading evidence. The medical and
economic consequences to EU member states and their residents/citizens could hardly be
higher.
IV. Furthermore, if the vaccines are approved without an appropriate and accurate review of
efficacy, then any potential acceptance or mandate of these vaccines is likely to be based on
inaccurate evidence regarding the vaccine, namely that it will stop transmission of the virus
from the vaccine recipient to others and/or that it will reduce COVID-19 disease and deaths.
The Pfizer/BioNTech trial protocol and other trial protocols are currently not designed to
determine whether either of those objectives can be met; and even if it was, if cases cannot be
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reliably identified, neither objective could be reliably met.
V. The public interest also weighs strongly in favor of the requested relief because
improving the accurate determination of primary endpoints (i) will comport with the best
scientific practices, (ii) increase public confidence in the efficacy of a product likely to be
mandated or intended for widespread use, and (iii) not doing so will have the opposite result
and create uncertainties regarding the efficacy of and need for the COVID-19 vaccines.
VI. Petitioner hereby incorporates the grounds, facts, arguments and opinions stated in the
“PETITION FOR ADMINISTRATIVE ACTION REGARDING CONFIRMATION OF
EFFICACY END POINTS OF THE PHASE III CLINICAL TRIALS OF COVID-19
VACCINES” which has been submitted to the FDA by Dr. Sin Hang Lee via electronic filing
on November 25, 2020 (Exhibit A - Docket No. FDA-2020-P-2225). Exhibit A attached hereto
shall be incorporated herein and shall be understood to be a part hereof as though included in
the body of this petition.
VII. Petitioner hereby also incorporates the grounds, facts, arguments and opinions stated in
the external peer review of the “Drosten-Test” (Exhibit B). Design flaws of certain RT-qPCR
tests that are identical to or modeled after what is sometimes called the “Drosten-Test” can lead
to false-positive results in trials designed such that PCR results are the primary evidence of
infection. Exhibit B attached hereto shall be incorporated herein and shall be understood to be a
part hereof as though included in the body of this petition.
VIII. For a vaccine to work, our immune system needs to be stimulated to produce a
neutralizing antibody, as opposed to a non-neutralizing antibody. A neutralizing antibody is one
that can recognize and bind to some region (‘epitope’) of the virus, and that subsequently
results in the virus either not entering or replicating in your cells. A non-neutralizing antibody is
one that can bind to the virus, but for some reason, the antibody fails to neutralize the infectivity
of the virus. In some viruses, if a person harbors a non-neutralizing antibody to the virus, a
subsequent infection by the virus can cause that person to elicit a more severe reaction to the
virus due to the presence of the non-neutralizing antibody. This is not true for all viruses, only
particular ones. This is called Antibody Dependent Enhancement (ADE), and is a common
problem with Dengue Virus, Ebola Virus, HIV, RSV, and the family of coronaviruses. In fact,
this problem of ADE is a major reason why many previous vaccine trials for other
coronaviruses failed. Major safety concerns were observed in animal models. If ADE occurs in
an individual, their response to the virus can be worse than their response if they had never
developed an antibody in the first place. This can cause a hyperinflammatory response, a
cytokine storm, and a generally dysregulation of the immune system that allows the virus to
cause more damage to our lungs and other organs of our body. In addition, new cell types
throughout our body are now susceptible to viral infection due to the additional viral entry
pathway. There are many studies that demonstrate that ADE is a persistent problem with
coronaviruses in general, and in particular, with SARS-related viruses. ADE has proven to be a
serious challenge with coronavirus vaccines, and this is the primary reason many of such
vaccines have failed in early in-vitro or animal trials. For example, rhesus macaques who were
vaccinated with the Spike protein of the SARS-CoV virus demonstrated severe acute lung
injury when challenged with SARS-CoV, while monkeys who were not vaccinated did not.
Similarly, mice who were immunized with one of four different SARS-CoV vaccines showed
histopathological changes in the lungs with eosinophil infiltration after being challenged with
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SARS-CoV virus.
IX. There are some concerning issues with the trial designs, spelled out by Dr. Peter Doshi in
the British Medical Journal. Dr. Doshi focuses on the two biggest issues. First, none of the
leading vaccine candidate trials is designed to test if the vaccine can reduce severe COVID-19
symptoms, defined as: hospital admissions, ICU or death. And, second, the trials are not
designed to test if the vaccine can interrupt transmission
(https://www.bmj.com/content/bmj/371/bmj.m4037.full.pdf). If neither of these conditions is
met, the vaccine in essence performs like a therapeutic drug, except a vaccine would be taken
prophylactically, even by the perfectly healthy, and more than likely carries a higher risk of
injury than a therapeutic drug. If this were to be true, then therapeutic drugs would be superior
to any COVID vaccine.
X. In the Pfizer/BioNTech mRNA vaccine candidate, polyethylene glycol (PEG) is found in

the fatty lipid nanoparticle coating around the mRNA. Seventy percent of people make
antibodies to PEG and most do not know it, creating a concerning situation where many could
have allergic, potentially deadly, reactions to a PEG-containing vaccine. PEG antibodies may
also reduce vaccine effectiveness. Pfizer/BioNTech is also inserting an ingredient derived from
a marine invertebrate, mNeonGreen, into its vaccine. The ingredient has bioluminescent
qualities, making it attractive for medical imaging purposes, but it is unclear why an injected
vaccine would need to have that quality. mNeonGreen has unknown antigenicity.
XI. Several vaccine candidates are expected to induce the formation of humoral antibodies
against spike proteins of SARS-CoV-2. Syncytin-1 (see Gallaher, B., “Response to nCoV2019
Against Backdrop of Endogenous Retroviruses” - http://virological.org/t/response-to-ncov2019-
against-backdrop-of-endogenous-retroviruses/396), which is derived from human endogenous
retroviruses (HERV) and is responsible for the development of a placenta in mammals and
humans and is therefore an essential prerequisite for a successful pregnancy, is also found in
homologous form in the spike proteins of SARS viruses. There is no indication whether
antibodies against spike proteins of SARS viruses would also act like anti-Syncytin-1
antibodies. However, if this were to be the case this would then also prevent the formation of a
placenta which would result in vaccinated women essentially becoming infertile. To my
knowledge, Pfizer/BioNTech has yet to release any samples of written materials provided to
patients, so it is unclear what, if any, information regarding (potential) fertility-specific risks
caused by antibodies is included.
According to section 10.4.2 of the Pfizer/BioNTech trial protocol, a woman of childbearing

potential (WOCBP) is eligible to participate if she is not pregnant or breastfeeding, and is using
an acceptable contraceptive method as described in the trial protocol during the intervention
period (for a minimum of 28 days after the last dose of study intervention).
This means that it could take a relatively long time before a noticeable number of cases of post-
vaccination infertility could be observed.
XII. It appears that Pfizer/BioNTech have not yet released any samples of written materials
provided to patients, so it is unclear what, if any, instructions/information patients/subjects were
given regarding ADE and PEG-related issues and (potential) fertility- or pregnancy-specific
issues.
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D. STAY URGENTLY REQUIRED
I. Petitioner any many EU residents/citizens will suffer irreparable harm because once the
EMA approves the COVID- 19 vaccine(s) in question, both governments of EU member states
and employers in the EU are most likely going to recommend them for widespread use, and
hence without the EMA assuring proper safety trials of the vaccines now, the Petitioner and
others will not have the opportunity to object to receiving the vaccine based on deficient
clinical trials later.
II. Furthermore, if the vaccines are licensed without an appropriate efficacy review and
without improving the accurate determination of primary endpoints, then any potential
acceptance or mandate of these vaccines are likely to be based on inaccurate beliefs and data
about the vaccines, namely that they will or might stop transmission of the virus from the
vaccine recipient to others and/or that it will reduce severe COVID-19 disease and deaths. The
trial protocols in question are not currently properly designed to determine whether either of
those objectives can be met.
III. This petition is also not frivolous and is being pursued in good faith as it seeks to
increase the scientific integrity and reliability of the trials of the COVID-19 vaccines.
IV. Finally, the public interest also weighs strongly in favor of the requested relief because
improving the accurate determination of primary endpoints (i) will comport with the best
scientific practices, (ii) increase public confidence in the efficacy of a vaccine expected to be
mandated or strongly recommended for widespread use, and (iii) not doing so will have the
opposite result in that it will create uncertainties regarding the efficacy of and need for the
COVID-19 vaccines.
V. The Petitioner therefore respectfully urges that this request be granted forthwith.
Respectfully submitted on my behalf and on behalf of Co-Petitioner Dr. Michael Yeadon:
__________________________________________________________
Dr. med. Wolfgang Wodarg
Exhibit A
Exhibit B
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VIA ELECTRONIC FILING
November 25, 2020
Division of Dockets Management

Department of Health and Human Services
Food and Drug Administration
Commissioner Stephen M. Hahn, M.D.
5630 Fishers Lane
Rm. 1061
Rockville, MD 20852
UNITED STATES DEPARTMENT OF HEALTH AND HUMAN SERVICES

AND THE FOOD AND DRUG ADMINISTRATION
PETITION FOR ADMINISTRATIVE :

ACTION REGARDING :
CONFIRMATION OF EFFICACY :
END POINTS OF THE PHASE III : Docket No. FDA-2020-P-2225
CLINICAL TRIALS OF COVID-19 :
VACCINES :
ADMINISTRATIVE STAY OF ACTION
This petition for a stay of action is submitted on behalf of Dr. Sin Hang Lee ( Petitioner )
pursuant to 21 C.F.R. § 10.35 and related and relevant provisions of the Federal Food, Drug, and
Cosmetic Act or the Public Health Service Act to request the Commissioner of Food and Drugs
(the Commissioner ) stay the Phase III trials of BNT162b (NCT04368728) to conform with the
requests in the Action Requested section below.
Because of the compelling need to ensure the safety and efficacy of any COVID-19 vaccine
licensed by the FDA, and to allow Petitioner the opportunity to seek emergency judicial relief
should the Commissioner deny its Petition, Petitioner respectfully requests that FDA act on the
instant Petition by December 11, 2020.
A. DECISION INVOLVED
1. Approval of trial design for Phase III trial of BNT162 (NCT04368728)1
1
NCT04368728 available at https://www.clinicaltrials.gov/ct2/show/NCT04368728 (last visited November 3, 2020).
1
B. ACTION REQUESTED
2. Stay the Phase III trial of BNT162 (NCT04368728) until its study design is
amended to provide that:
Before an EUA or unrestricted license is issued for the Pfizer

vaccine, or for other vaccines for which PCR results are the primary
evidence of infection, all endpoints or COVID-19 cases used to
determine vaccine efficacy in the Phase 3 or 2/3 trials should have
their infection status confirmed by Sanger sequencing, given the
high cycle thresholds used in some trials. High cycle thresholds, or
Ct values, in RT-qPCR test results have been widely acknowledged
to lead to false positives.2
All RT-qPCR-positive test results used to categorize patient as

COVID-19 cases and used to qualify the trial s endpoints should
be verified by Sanger sequencing to confirm that the tested samples
in fact contain a unique SARS-CoV-2 genomic RNA. Congruent
with FDA requirements for a confirmed diagnosis of human
papillomavirus (HPV) using PCR, the sequencing electropherogram
must show a minimum of 100 contiguous bases matching the
reference sequence with an Expected Value (E Value) <10-30 for the
specific SARS-CoV-2 gene sequence based on a BLAST search of
the GenBank database (aka NCBI Nucleotide database).
C. STATEMENT OF GROUNDS
3. As detailed herein, (i) without the requested stay, the Petitioner will suffer
irreparable harm, (ii) the request is not frivolous and is being pursued in good faith, (iii) the request
demonstrates sound public policy, and (iv) the public interest favors granting a stay. 3
4. The current study designs for the Phase II/III trials of BNT162b ( the Pfizer
Vaccine ) are inadequate to accurately assess efficacy.
5. Petitioner and the public will suffer irreparable harm if the actions requested herein
are not granted, because once the FDA licenses this COVID-19 vaccine, both governments and
employers may make this product mandatory (in general, or for airline or international travel) or
may recommend it for widespread use. If the assignment of cases and non-cases during the course
of the trial is not accurate, the vaccine will not have been properly tested. If the vaccine is not
2
See New York Times. Your Coronavirus Test Is Positive. Maybe It Shouldn t Be. By Apoorva Mandavilli. Published
Aug. 29, 2020 and updated Sept. 17, 2020, available at https://www.nytimes.com/2020/08/29/health/coronavirus-
testing.html.
3
The Petitioner hereby incorporates by reference as if fully set forth herein the Statement of Grounds from its Citizen s
Petition, dated November 23, 2020, available at, https://beta.regulations.gov/document/FDA-2020-P-2225 (last
visited November 25, 2020).
2
properly tested, important public policy decisions regarding its use will be based on misleading
evidence. The medical and economic consequences to the nation could hardly be higher.
6. The New York State Bar Association has already issued a report on COVID-19
recommending that, a vaccine subject to scientific evidence of safety and efficacy be made widely
available, and widely encouraged, and if the public health authorities conclude necessary,
4
required Thus, it is reasonable to suspect that COVID-19 vaccines, including the Pfizer
vaccine, could become mandatory. Without the FDA assuring proper efficacy trials of the vaccine
now, the Petitioner and the public may not have the opportunity to object to receiving the vaccine,
which was approved based on currently deficient and unreliable clinical trial data.
7. Furthermore, if the vaccine is approved without an appropriate and accurate review

of efficacy, then any potential acceptance or mandate of these vaccines is likely to be based on
inaccurate evidence regarding the vaccine, namely that it will stop transmission of the virus from
the vaccine recipient to others and/or that it will reduce severe COVID-19 disease and deaths. The
Pfizer trial protocol is currently not designed to determine whether either of those objectives can
be met; and even if it was, if cases cannot be reliably identified, neither objective could be reliably
met.
8. The public interest also weighs strongly in favor of the requested relief because
improving the accurate determination of primary endpoints (i) will comport with the best scientific
practices, (ii) increase public confidence in the efficacy of a product likely to be mandated or
intended for widespread use, and (iii) not doing so will have the opposite result and create
uncertainties regarding the efficacy of and need for the COVID-19 vaccines.
7. According to the trial protocol, 8.1. Efficacy and/or Immunogenicity

Assessments, the trial s primary endpoint is prevention of symptomatic disease in vaccine
recipients. In order to evaluate that endpoint, the trial will track recorded COVID-19 disease. The
definition of confirmed COVID-19 is:
presence of at least 1 of the following symptoms and SARS-CoV-2 NAAT-positive

during, or within 4 days before or after, the symptomatic period, either at the central
laboratory or at a local testing facility (using an acceptable test):
Fever;
New or increased cough;
New or increased shortness of breath;
Chills;
New or increased muscle pain;
New loss of taste or smell;
Sore throat;
Diarrhea;
Vomiting.
4
https://nysba.org/app/uploads/2020/06/2b-REV-6-12-20-FINAL-HOD-RESOLUTIONS-1-through-4.pdf.
3
8. As a result, if a participant has a positive reverse transcription-quantitative
polymerase chain reaction ( RT-qPCR ) test along with a cough or sore throat, that participant
would be considered as a confirmed COVID-19 case and would be counted as an endpoint.
Once a trial reaches a certain number of endpoints , the trial is closer to seeking FDA approval
or licensure by demonstrating that the vaccine is effective (in that the vaccine group had lower
incidence of endpoints than the control group).
9. This effectively means that the efficacy of the vaccine will be determined based on
only symptoms of non-specific disease in conjunction with a PCR positive laboratory test.
10. According to the trial protocol, 8.1 Efficacy and/or Immunogenicity

Assessments, efficacy will be assessed throughout a participant s involvement in the study
through surveillance for potential cases of COVID-19. If, at any time, a participant develops acute
respiratory illness (see Section 8.13), for the purposes of the study he or she will be considered to
potentially have COVID-19 illness. In this circumstance, the participant should contact the site, an
in-person or telehealth visit should occur, and assessments should be conducted as specified in the
SoA. The assessments will include a nasal (midturbinate) swab, which will be tested at a central
laboratory using a reverse transcription polymerase chain reaction (RT-PCR) test (Cepheid; FDA
approved under EUA), or other equivalent nucleic acid amplification based test (ie, NAAT), to
detect SARS-CoV-2. In addition, clinical information and results from local standard-of-care tests
(as detailed in Section 8.13) will be assessed. The central laboratory NAAT result will be used for
the case definition, unless no result is available from the central laboratory, in which case a local
NAAT result may be used if it was obtained using 1 of the following assays:
Cepheid Xpert Xpress SARS-CoV-2

Roche cobas SARS-CoV-2 real-time RT-PCR test
(EUA200009/A001)
Abbott Molecular/RealTime SARS-CoV-2 assay
(EUA200023/A001)
11. These test kits referred to in the trial protocol, namely the Cepheid Xpert Xpress
SARS-CoV-2, the Roche cobas SARS-CoV-2 real-time RT-PCR test (EUA200009/A001), and
the Abbott Molecular/RealTime SARS-CoV-2 assay (EUA200023/A001), are very unreliable
tools when they are used to determine whether the nasal swab sample collected from a
symptomatic participant contains SARS-CoV-2 or not. These real-time RT-PCR or RT-
quantitative PCR tests should be referred to as rRT-PCR or RT-qPCR tests to be distinguished
from conventional RT-PCR. The very short RT-qPCR product (amplicon) cannot be analyzed by
automated Sanger sequencing as the products of conventional PCR can. And DNA sequencing for
validation of the PCR products is needed to correctly determine if the presumptive RT-qPCR-
positive SARS-CoV-2 test result is a true positive or a false positive. The reasoning is further
outlined as follows:
a. Nowadays DNA sequencing of the PCR amplicon of the genomic nucleic acid of the
pathogen is a universally accepted technology for detection and for confirmation of
infectious agents, especially pathogenic viruses, in clinical specimens. On January 10,
4
2020, the first SARS-CoV-2 genome sequence was released online. On the same day, a
group of American scientists, most from the CDC, immediately designed 2 complementary
panels of primers to amplify the virus genome for sequencing. The PCR amplicons
averaged 550 bp in size in their research.5
b. The World Health Organization (WHO) guidance titled WHO Laboratory testing for
coronavirus disease (COVID-19) in suspected human cases-Interim guidance dated 19
March 2020 advised Routine confirmation of cases of COVID-19 is based on detection
of unique sequences of virus RNA by NAAT such as real-time reverse transcription-
polymerase chain reaction (rRT-PCR) with confirmation by nucleic acid sequencing when
necessary. 6
c. The FDA also recognizes the inherent inaccuracy of the RT-qPCR tests. In its letter issued
on February 4, 2020 authorizing emergency use of the CDC 2019-Novel Coronavirus
(2019-nCoV, renamed as SARS-CoV-2) Real-Time Reverse Transcriptase (RT)-PCR
Diagnostic Panel, the FDA specifically stated that the test panel is for the presumptive
qualitative detection of nucleic acid from the 2019-nCoV (sic) in upper and lower
respiratory specimens. 7
d. In addition to false-negative results, these RT-qPCR test kits under EUA also generate
false-positive test results. For example, 77 positive SARS-CoV-2 test results on a group
of football players all turned out to be false positives on repeat tests.8
e. The FDA has officially alerted clinical laboratory staff and health care providers of an
increased risk of false-positive results with some of these commercial test kits permitted to
be used under EUA.9
5
Paden CR, Tao Y, Queen K, Zhang J, Li Y, Uehara A, Tong S. Rapid, Sensitive, Full-Genome Sequencing of Severe
Acute Respiratory Syndrome Coronavirus 2. Emerg Infect Dis. 2020 Oct;26(10):2401-2405. doi:
10.3201/eid2610.201800. Epub 2020 Jul 1. PMID: 32610037; PMCID: PMC7510745.
6
WHO Laboratory testing for coronavirus disease (COVID-19) in suspected human cases-Interim guidance 19 March
2020. Available from: https://www.who.int/publications/i/item/10665-331501.
7
FDA letter dated February 4, 2020 authorizing emergency use of the CDC 2019-Novel Coronavirus (2019-nCoV,
renamed as SARS-CoV-2) Real-Time Reverse Transcriptase (RT)-PCR Diagnostic Panel. See Open letter from FDA
to Robert R. Redfield, MD, Director, Centers for Disease Control and Prevention. March 15, 2020.
https://www.fda.gov/media/134919/download.
8
Kevin Patra. Around the NFL- All 77 false-positive COVID-19 tests come back negative upon reruns. Aug 24, 2020.
Available from: https://www.nfl.com/news/all-77-false-positive-covid-19-tests-come-back-negative-upon-reruns.
9
FDA. False Positive Results with BD SARS-CoV-2 Reagents for the BD Max System - Letter to Clinical Laboratory
Staff and Health Care Providers. Available from: https://www.fda.gov/medical-devices/letters-health-care-
providers/false-positive-results-bd-sars-cov-2-reagents-bd-max-system-letter-clinical-laboratory-staff-and Accessed
November 2, 2020; see also FDA. Risk of Inaccurate Results with Thermo Fisher Scientific TaqPath COVID-19
Combo Kit - Letter to Clinical Laboratory Staff and Health Care Providers. Available from:
https://www.fda.gov/medical-devices/letters-health-care-providers/risk-inaccurate-results-thermo-fisher-scientific-
taqpath-covid-19-combo-kit-letter-clinical?utm_campaign=2020-08-17%20Risk%20of%20Inaccurate%20Results
%20with%20Thermo%20Fisher%20Scientific%20TaqPath&utm_medium=email&utm_source=Eloqua.
5
f. To resolve the problems caused by these inherently inaccurate tests, the FDA s position is
that false results can be investigated using an additional EUA RT-qPCR assay, and/or
Sanger sequencing.10 Since an additional EUA RT-qPCR test result may also generate a
false result, Sanger sequencing is the de facto gold standard for confirmation of
presumptive qualitative detection of nucleic acid from the SARS-CoV-2 and for excluding
false-positive cases.
g. According to the FDA guidance on molecular diagnosis of viral infection caused by human
papillomavirus (HPV), a conventional PCR detection of genomic DNA followed by Sanger
sequencing on both strands of the PCR amplicon (bi-directional sequencing) that contains
a minimum of 100 contiguous bases is acceptable as valid diagnostics for HPV infection
provided the sequence matches the reference or consensus sequence, e.g. with an Expected
Value (E Value) <10-30 for the specific HPV DNA target based on a BLAST search of the
GenBank (NCBI Nucleotide) database.11 Following this FDA guidance, and showing the
feasibility of implementing the FDA guidance for accurate diagnosis of COVID-19, a
protocol using the nested PCR cDNA amplicon of a 398-base highly conserved SARS-
CoV-2 N gene segment as the template for Sanger sequencing was developed for
confirmatory detection of SARS-CoV-2 in clinical samples.12
h. DNA sequencing verification is necessary for confirmation of the presumptive SARS-

CoV-2-positive cases in the Pfizer vaccine s Phase II/III clinical trial because, according
to its Protocol, the specimens collected from the symptomatic trial subjects were sent to a
central laboratory using a reverse transcription polymerase chain reaction (RT-PCR) test
(Cepheid; FDA approved under EUA), or other equivalent nucleic acid amplification
based test (i.e., NAAT), to detect SARS-CoV-2.
In order to raise the detection sensitivity, the mean Ct value of the Cepheid system is set
as high as 42.9 for the N2 target, and as high as 44.9 for the E target, as shown in Table 4
of Instructions for Users (Cepheid 302-3562, Rev. E September 2020).13
10
FDA. Molecular Diagnostic Template for Laboratories. Policy for Coronavirus Disease-2019 Tests During the
Public Health Emergency (Revised) Available from: https://www.fda.gov/media/135659/download .
11
FDA. Establishing the Performance Characteristics of In Vitro Diagnostic Devices for the Detection or Detection
and Differentiation of Human Papillomaviruses. Available from: https://www.fda.gov/media/92930/download.
12
Lee SH. Testing for SARS-CoV-2 in cellular components by routine nested RT-PCR followed by DNA sequencing.
International Journal of Geriatrics and Rehabilitation. 2020; 2:69-96. Available from: http://www.int-soc-clin-
geriat.com/info/wpcontent/uploads/2020/03/Dr.-Lees-paper-on-testing-for-SARS-CoV-2.pdf.
13
Cepheid. GeneXpert. Instructions for Users. XPRSARS-COV2-10. 302-3562, Rev. E September 2020
https://www.cepheid.com/Package%20Insert%20Files/Xpress-SARS-CoV-2/Xpert%20Xpress%20SARS-CoV-
2%20Assay%20ENGLISH%20Package%20Insert%20302-3562-GX%20Rev.%20E.pdf.
6
At Ct values between 36.0 and 44.9, many RT-qPCR positive test results are false positives.
i. The results of the 3 RT-qPCR test kits used in the trial protocol are not comparable. A
sample identified as negative by the Abbott kit can be classified as positive by the Cepheid
kit. According to an FDA survey, the limit of detection by the Cepheid Xpert Xpress
SARS-CoV-2 test kit and the limit of detection by Abbott RealTime SARS-CoV-2 assay
kit are found to be identical, namely both being at 5400 NAAT Detectable Units/ mL, as
shown in the comparative data extracted from an FDA reference panel.14
5400 Cepheid Xpert Xpress SARS-CoV-2 test
5400 Abbott Molecular Abbott RealTime SARS-CoV-2 assay
However, due to the designation of higher cycle threshold test results as positives, the
Cepheid Xpert kits have classified many Abbott kit negative cases as positives in a head-
to-head comparative study as shown in the following Table 2 extracted from a report by
Basu et al.15
14
FDA. SARS-CoV-2 Reference Panel Comparative Data. https://www.fda.gov/medical-devices/coronavirus-covid-
19-and-medical-devices/sars-cov-2-reference-panel-comparative- data.
15
See bioRxiv preprint doi: https://doi.org/10.1101/2020.05.11.089896; Basu A, Zinger T, Inglima K, Woo KM, Atie
O, Yurasits L, See B, Aguero-Rosenfeld ME. Performance of Abbott ID Now COVID-19 Rapid Nucleic Acid
Amplification Test Using Nasopharyngeal Swabs Transported in Viral Transport Media and Dry Nasal Swabs in a
New York City Academic Institution. J Clin Microbiol. 2020 Jul 23;58(8):e01136-20. doi: 10.1128/JCM.01136-20.
PMID: 32471894; PMCID: PMC7383552.
7
j. One of the Cepheid Xpert kit users has put out an alert, stating The instruments are
presently set by the manufacturer to interpret a single target positive with very poor
amplification efficiency (high Cycle Threshold [Ct] and/or atypical curve) as
DETECTED. None of these to date have confirmed positive when tested on other
systems using similar targets, and may be a false positive due to background noise. 16
k. Another group of users also found that some tested samples classified as positives by the
Cepheid test kits cannot be confirmed with other test kits. These authors published a report,
stating: We found that the sensitivity of the Xpert Xpress SARS-CoV-2 assay was 100%
(20 of 20) and the specificity was 80% (16 of 20). When looking at the cycle threshold
(Ct) values from the GeneXpert assay we observed that specimens with no amplification
of the E gene (ie, Ct=0) and Ct values for the N2 gene greater than 40 cycles were
considered as positives, whereas they were negative using the other RT-PCR system (Da
An Gene). 17
16
Diagnostic Laboratory Services Inc. Technical Alert. Cepheid GeneXpert and BD Max Instruments may be
Reporting False Positives. https://dlslab.com/documents/bulletins/2020/tech-memo-sars-cov-2-pcr-possible-false-
positive-6-19-2020.pdf.
17
Rakotosamimanana N, Randrianirina F, Randremanana R, Raherison MS, Rasolofo V, Solofomalala GD, Spiegel
A, Heraud JM. GeneXpert for the diagnosis of COVID-19 in LMICs. Lancet Glob Health. 2020 Oct 19:S2214-
109X(20)30428-9. doi: 10.1016/S2214-109X(20)30428-9. Epub ahead of print. PMID: 33091372; PMCID:
PMC7572106.
8
12. DNA sequencing verification of the RT-qPCR positive test results is absolutely
necessary in this placebo-controlled randomized clinical trial because de facto unblinding has
occurred among the participants. According to the trial protocol Section 8.13. COVID-19
Surveillance (All Participants), If a participant experiences any of the following (irrespective of
perceived etiology or clinical significance), he or she is instructed to contact the site immediately
and, if confirmed, participate in an in-person or telehealth visit as soon as possible. This contact
would trigger an automatic NAAT test by a Cepheid RT-qPCR assay at the central laboratory or
at a local laboratory by any similar acceptable methods.
At the time of enrollment, the participants were informed that each of them would be
injected with a vaccine to protect against COVID-19 infection or a saline placebo without
disclosing which one of the two was injected into the participant. However, all participants were
also informed that the vaccine may cause the following reactions:
Fever 39.0°C ( 102.1°F).

Redness or swelling at the injection site measuring greater than 10
cm (>20 measuring device units).
Severe pain at the injection site.
Any severe systemic event.
It is commonly known to the general public and especially to the informed clinical trial
participants that intramuscular injection of a very small amount of sterile normal saline will not
cause fever, local redness and swelling, and severe pain, or systemic reactions. The participants
receiving placebo would intuitively or reasonably know that they were not injected with a vaccine
and were not protected against COVID-19 disease due to the lack of any vaccine reaction after the
injection. As a result, more participants receiving placebo than those receiving vaccine would
report to the site manager when they developed any minor symptoms, such as a sore throat or a
new cough for the fear of coming down with COVID-19. The site manager must investigate the
symptoms reported, including ordering a RT-qPCR test by Cepheid assay to be performed at the
Central Laboratory according to Protocol. The more severe cases might be tested locally by Abbott
kits or Roche kits because they might have to be tested in the hospital after admission, and because
many hospitals are aware of the high false positive rates generated by the Cepheid kits. The results
generated by these test kits are not comparable since the Cepheid test kits using a very high Ct
value up to 44.9 for detection of SARS-CoV-2 genomic RNA tend to generate many more
false positives than the other test kits. A higher number of false-positive test results in the
participants receiving placebo will artificially raise the efficacy of the vaccine, unless the RT-
qPCR test results are verified by nucleotide sequencing to eliminate all false-positive test results.
13. Based on an MPR report published on November 8, 2020, there are only 180
confirmed cases of COVID-19 in this clinical trial series that have been analyzed to support the
vaccine efficacy evaluation.18 If the Sponsor (BioNTech/Pfizer) is unable to perform confirmatory
Sanger sequencing tests on these 180 RNA extract residual samples, the Petitioner hereby offers
18
Diana Ernst, RPh. Final Analysis Reveals COVID-19 Vaccine Candidate BNT162b2 95% Effective. MPR Report.
November 18, 2020. https://www.empr.com/home/news/drugs-in-the-pipeline/pfizer-biontech-mrna-based-vaccine-
bnt162b2-against-covid19-effective/.
9
to re-test them immediately with Sanger sequencing19 and submit the laboratory data to support
FDA s evaluation. Therefore, there is no excuse for the Sponsor to refuse using the gold standard
Sanger sequencing technology for endpoint validation.
14. In summary, based on the scientific data available in the public domain and the
FDA guidance, all RT-qPCR test results for detection of SARS-CoV-2 gene sequence must be
considered presumptive. The Cepheid test kits for SARS-CoV-2 are known to generate more false-
positive test results than other EUA assay kits.
15. The residues of the tested samples that were classified as positive for SARS-CoV-
2 by the Cepheid GeneXpert assay, or equivalent as stated in the Pfizer Clinical Trial Protocol,
must be re-tested by a Sanger sequencing method to confirm that the presumptive positive samples
in fact contain a unique sequence of SARS-CoV-2 genome. Only then can the positive test results
from the Cepheid GeneXpert test kits be accepted as an accurate component of the endpoint.
Only then can one nonspecific symptom plus laboratory positivity be accepted as a valid measure
of confirmed COVID-19 cases or endpoints.
Stay Urgently Required
16. Petitioner will suffer irreparable harm because once the FDA licenses this COVID-
19 vaccine, states are expected to make this product mandatory, and hence without the FDA
assuring proper safety trials of the vaccine now, the Petitioner will not have the opportunity to
object to receiving the vaccine based on deficient clinical trials later.
17. For example, the New York State Bar Association recently passed a resolution
recommending that [s]hould the level of immunity be deemed insufficient by expert medical and
scientific consensus to check the spread of COVID-19 and reduce morbidity and mortality, a
mandate and state action should be considered. 20 Mandating administration of the vaccine,
thereby eliminating the right to informed consent, makes acute the need to assure that the safety
and efficacy of any COVID-19 vaccine is robustly studied in an adequate clinical trial monitoring
for any potential adverse events.
18. Furthermore, if the vaccine is licensed without an appropriate efficacy review and
without improving the accurate determination of primary endpoints, then any potential acceptance
or mandate of these vaccines are likely to be based on inaccurate beliefs about the vaccine, namely
that it will stop transmission of the virus from the vaccine recipient to others or that it will reduce
severe COVID-19 disease and deaths. The trial protocols are not currently designed to determine
whether either of those objectives can be met.
19
Lee SH. Testing for SARS-CoV-2 in cellular components by routine nested RT-PCR followed by DNA sequencing.
International Journal of Geriatrics and Rehabilitation. 2020; 2:69-96. Available from: http://www.int-soc-clin-
geriat.com/info/wpcontent/uploads/2020/03/Dr.-Lees-paper-on-testing-for-SARS-CoV-2.pdf.
20
https://nysba.org/app/uploads/2020/11/11.-Health-Law-Section-COVID-19-Report-September-20-2020-with-all-
comments.pdf (emphasis added) (last visited November 10, 2020).
10
19. This request is also not frivolous and is being pursued in good faith as it seeks to
increase the scientific integrity and reliability of the trials of the COVID-19 Vaccines.
20. Finally, the public interest also weighs strongly in favor of the requested relief
because improving the accurate determination of primary endpoints (i) will comport with the best
scientific practices, (ii) increase public confidence in the efficacy of a product expected to be
mandated, and (iii) not doing so will have the opposite result in that it will create uncertainties
regarding the efficacy of and need for the COVID-19 Vaccines.
21. The Petitioner therefore respectfully urges that this request be granted forthwith.
Respectfully submitted,
Dr. Sin Hang Lee
11
Review Report by an International Consortium of Scientists in Life Sciences ICSLS
Corman Drosten e al , Eurosurveillance Updated: . .
External peer review of the RTPCR test to detect SARS-CoV-

reveals major scientific flaws at the molecular and
methodological level: consequences for false positive results.
Pieter Borger , Bobby Rajesh Malhotra , Michael Yeadon , Clare Craig
Kevin McKernan , Klaus Steger , Paul McSheehy , Lidiya Angelova
Fabio Franchi , Thomas Binder , Henrik Ullrich
, Makoto Ohashi
Stefano Scoglio , Marjolein Doesburg van Kleffens , Dorothea Gilbert
Rainer Klement , Ruth Schruefer , Berber W. Pieksma , Jan Bonte
Bruno H. Dalle Carbonare , Kevin P. Corbett , Ulrike Kämmerer
Corresponding Author
ABSTRACT
In the publication entitled Detection of novel coronavirus nCoV by real time RT PCR
Eurosurveillance the authors present a diagnostic workflow and RT qPCR protocol for
detection and diagnostics of nCoV now known as SARS CoV , which they claim to be
validated, as well as being a robust diagnostic methodology for use in public health laboratory
settings.
In light of all the consequences resulting from this very publication for societies worldwide, a group
of independent researchers performed a point by point review of the aforesaid publication in which
all components of the presented test design were cross checked, the RT qPCR
protocol recommendations were assessed with respect to good laboratory practice, and
parameters examined against relevant scientific literature covering the field.
The published RT qPCR protocol for detection and diagnostics of nCoV and the manuscript
suffer from numerous technical and scientific errors, including insufficient primer design, a
problematic and insufficient RT qPCR protocol, and the absence of an accurate test validation.
Neither the presented test nor the manuscript itself fulfils the requirements for an acceptable
scientific publication. Further, serious conflicts of interest of the authors are not mentioned. Finally,
the very short timescale between submission and acceptance of the publication hours signifies
that a systematic peer review process was either not performed here, or of problematic poor quality.
We provide compelling evidence of several scientific inadequacies, errors and flaws. Considering the
scientific and methodological blemishes presented here, we are confident that the editorial board of
Eurosurveillance has no other choice but to retract the publication.
CONCISE REVIEW REPORT

This paper will show numerous serious flaws in the Corman Drosten paper, the significance of which
has led to worldwide misdiagnosis of infections attributed to SARS CoV and associated with the
disease COVID . We are confronted with stringent lockdowns which have destroyed many people’s
lives and livelihoods, limited access to education and these imposed restrictions by governments
around the world are a direct attack on people’s basic rights and their personal freedoms, resulting in
collateral damage for entire economies on a global scale.
There are ten fatal problems with the Corman-Drosten paper which we will outline and explain in
greater detail in the following sections.
The first and major issue is that the novel Coronavirus SARS CoV in the publication named
nCoV and in February named SARS CoV by an international consortium of virus experts
is based on in silico theoretical sequences, supplied by a laboratory in China , because at the time
neither control material of infectious live or inactivated SARS CoV nor isolated genomic RNA of
the virus was available to the authors. To date no validation has been performed by the authorship
based on isolated SARS CoV viruses or full length RNA thereof. According to Corman et al.:
We aimed o de elo and de lo ob diagno ic me hodolog fo e in blic

heal h labo a o e ing i ho ha ing i ma e ial a ailable
The focus here should be placed upon the two stated aims: a de elo men and b de lo men of a
diagno ic e fo e in blic heal h labo a o e ing . These aims are not achievable without
having any actual virus material available e.g. for determining the infectious viral load . In any case,
only a protocol with maximal accuracy can be the mandatory and primary goal in any
scenario outcome of this magnitude. Critical viral load determination is mandatory information, and
it is in Christian Drosten’s group responsibility to perform these experiments and provide the crucial
data.
Nevertheless these in silico sequences were used to develop a RT PCR test methodology to identify
the aforesaid virus. This model was based on the assumption that the novel virus is very similar to
SARS CoV from as both are beta coronaviruses.
The PCR test was therefore designed using the genomic sequence of SARS CoV as a control material
for the Sarbeco component; we know this from our personal email communication with one of
the co authors of the Corman Drosten paper. This method to model SARS CoV was described in the
Corman Drosten paper as follows:
he e abli hmen and alida ion of a diagno ic o kflo fo nCoV c eening

and ecific confi ma ion de igned in ab ence of a ailable i i ola e o o iginal
a ien ecimen De ign and alida ion e e enabled b he clo e gene ic ela edne
o he SARS CoV and aided b he e of n he ic n cleic acid echnolog
The Reverse Transcription Polymerase Chain Reaction RT PCR is an important biomolecular

technology to rapidly detect rare RNA fragments, which are known in advance. In the first step, RNA
molecules present in the sample are reverse transcribed to yield cDNA. The cDNA is then amplified in
the polymerase chain reaction using a specific primer pair and a thermostable DNA polymerase
enzyme. The technology is highly sensitive and its detection limit is theoretically molecule of cDNA.
The specificity of the PCR is highly influenced by biomolecular design errors.
What is important when designing an RT-PCR Test and the quantitative RT-qPCR
test described in the Corman-Drosten publication?
. The primers and probes:
a the concentration of primers and probes must be of optimal range nM
b must be specific to the target gene you want to amplify
c must have an optimal percentage of GC content relative to the total nitrogenous bases minimum
, maximum
d for virus diagnostics at least primer pairs must detect viral genes preferably as far apart as
possible in the viral genome
. The temperature at which all reactions take place:
a DNA melting temperature
b DNA amplification temperature TaqPol specific
c Tm; the annealing temperature the temperature at which the primers and probes reach the target
binding detachment, not to exceed ̊C per primer pair . Tm heavily depends on GC content of the
primers
. The number of amplification cycles less than ; preferably - cycles ;
In case of virus detection, cycles only detects signals which do not correlate with infectious virus
as determined by isolation in cell culture reviewed in ; if someone is tested by PCR as positive
when a threshold of cycles or higher is used as is the case in most laboratories in Europe the
US , the probability that said person is actually infected is less than , the probability that said
result is a false positive is reviewed in
. Molecular biological validations; amplified PCR products must be validated either by running the products
in a gel with a DNA ruler, or by direct DNA sequencing
. Positive and negative controls should be specified to confirm/refute specific virus detection
. There should be a Standard Operational Procedure SOP available
SOP unequivocally specifies the above parameters, so that all laboratories are able to set up the
exact same test conditions. To have a validated universal SOP is essential, because it enables the
comparison of data within and between countries.
MINOR CONCERNS WITH THE CORMAN-DROSTEN PAPER
. In Table of the Corman Drosten paper, different abbreviations are stated nM is specified,
nm isn’t. Further in regards to correct nomenclature, nm means nanometer therefore nm should
read nM here.
. It is the general consensus to write genetic sequences always in the ’ ’ direction, including the
reverse primers. It is highly unusual to do alignment with reverse complementary writing of the
primer sequence as the authors did in figure of the Corman Drosten paper. Here, in addition, a
wobble base is marked as y without description of the bases the Y stands for.
. Two misleading pitfalls in the Corman Drosten paper are that their Table does not include
Tm values annealing temperature values , neither does it show GC values number of G and C in the
sequences as value of total bases .
MAJOR CONCERNS WITH THE CORMAN-DROSTEN PAPER
A BACKGROUND
The authors introduce the background for their scientific work as: The ongoing outbreak of the
recently emerged novel coronavirus nCoV poses a challenge for public health laboratories as
virus isolates are unavailable while there is growing evidence that the outbreak is more widespread
than initially thought, and international spread through travelers does already occur .
According to BBC News and Google Statistics there were deaths world wide on January st
the day when the manuscript was submitted. Why did the authors assume a challenge for
public health laboratories while there was no substantial evidence at that time to indicate that the
outbreak was more widespread than initially thought?
As an aim the authors declared to develop and deploy robust diagnostic methodology for use in
public health laboratory settings without having virus material available. Further, they acknowledge
that The present study demonstrates the enormous response capacity achieved through
coordination of academic and public laboratories in national and European research networks.
B METHODS AND RESULTS
. Primer Probe Design
a Erroneous primer concentrations

Reliable and accurate PCR test protocols are normally designed using between nM and nM
per primer . In the Corman Drosten paper, we observe unusually high and varying primer
concentrations for several primers table . For the RdRp SARSr F and RdRp SARSr R primer pairs,
nM and nM are described, respectively. Similarly, for the N Sarbeco F and N Sarbeco R
primer set, they advise nM and nM, respectively .
It should be clear that these concentrations are far too high to be optimal for specific amplifications
of target genes. There exists no specified reason to use these extremely high concentrations of
primers in this protocol. Rather, these concentrations lead to increased unspecific binding and PCR
product amplification.
Table : Primers and probes adapted from Corman Drosten paper; erroneous primer concentrations are
highlighted
b Unspecified “Wobbly primer and probe sequences

To obtain reproducible and comparable results, it is essential to distinctively define the
primer pairs. In the Corman Drosten paper we observed six unspecified positions, indicated
by the letters R, W, M and S Table . The letter W means that at this position there can be
either an A or a T; R signifies there can be either a G or an A; M indicates that the position

may either be an A or a C; the letter S indicates there can be either a G or a C on this
position. This high number of variants not only is unusual, but it also is highly confusing for
laboratories. These six unspecified positions could easily result in the design of several
different alternative primer sequences which do not relate to SARS CoV distinct
RdRp SARSr F primers distinct RdRp SARS P probes distinct RdRp SARSr R . The
design variations will inevitably lead to results that are not even SARS CoV related.
Therefore, the confusing unspecific description in the Corman Drosten paper is not suitable
as a Standard Operational Protocol. These unspecified positions should have been designed
unequivocally.
These wobbly sequences have already created a source of concern in the field and resulted
in a Letter to the Editor authored by Pillonel et al. regarding blatant errors in the
described sequences. These errors are self evident in the Corman et al. supplement as well.
Table : Primers and probes adapted from Corman Drosten paper; unspecified Wobbly nucleotides in the
primers are highlighted
The WHO protocol Figure , which directly derives from the Corman Drosten paper,
concludes that in order to confirm the presence of SARS CoV , two control genes the
E and the RdRp genes must be identified in the assay. It should be noted, that the
RdPd gene has one uncertain position wobbly in the forward primer R G A , two
uncertain positions in the reverse primer R G A; S G C and it has three uncertain
positions in the RdRp probe W A T; R G A; M A C . So, two different forward primers,
four different reverse primers, and eight distinct probes can be synthesized for the
RdPd gene. Together, there are possible combinations of primers and probes!
The Corman Drosten paper further identifies a third gene which, according to the WHO
protocol, was not further validated and deemed unnecessary:
Of no e he N gene a a al o e fo med ell b a no bjec ed

o in en i e f he alida ion beca e i a ligh l le en i i e
This was an unfortunate omission as it would be best to use all three gene PCRs as
confirmatory assays, and this would have resulted in an almost sufficient virus RNA
detection diagnostic tool protocol. Three confirmatory assay steps would at least
minimize out errors uncertainties at every fold step in regards to Wobbly spots.
Nonetheless, the protocol would still fall short of any good laboratory practice , when
factoring in all the other design errors .
As it stands, the N gene assay is regrettably neither proposed in the WHO recommendation
Figure as a mandatory and crucial third confirmatory step, nor is it emphasized in the
Corman Drosten paper as important optional reassurance for a routine workflow Table .
Consequently, in nearly all test procedures worldwide, merely primer matches were used
instead of all three. This oversight renders the entire test protocol useless with regards to
delivering accurate test results of real significance in an ongoing pandemic.
Figure : The N Gene confirmatory assay is neither emphasized as necessary third step in the official WHO
Drosten Corman protocol recommendation below nor is it required as a crucial step for higher test accuracy
in the Eurosurveillance publication.
c Erroneous GC content discussed in c, together with annealing temperature Tm
d Detection of viral genes

RT PCR is not recommended for primary diagnostics of infection. This is why the RT PCR Test
used in clinical routine for detection of COVID is not indicated for COVID diagnosis on
a regulatory basis.
Clinician need o ecogni e he enhanced acc ac and eed of he molec la diagno ic

echni e fo he diagno i of infec ion b al o o nde and hei limi a ion Labo a o
e l ho ld al a be in e e ed in he con e of he clinical e en a ion of he a ien
and a o ia e i e ali and iming of ecimen collec ion a e e i ed fo eliable e
e l
However, it may be used to help the physician’s differential diagnosis when he or she has to
discriminate between different infections of the lung Flu, Covid and SARS have very
similar symptoms . For a confirmative diagnosis of a specific virus, at least specific primer
pairs must be applied to detect virus specific genes. Preferably, these target genes should
be located with the greatest distance possible in the viral genome opposite ends included .
Although the Corman Drosten paper describes primers, these primers only cover roughly
half of the virus’ genome. This is another factor that decreases specificity for detection of
intact COVID virus RNA and increases the quote of false positive test results.
Therefore, even if we obtain three positive signals i.e. the three primer pairs give different
amplification products in a sample, this does not prove the presence of a virus. A better
primer design would have terminal primers on both ends of the viral genome. This is
because the whole viral genome would be covered and three positive signals can better
discriminate between a complete and thus potentially infectious virus and fragmented viral
genomes without infectious potency . In order to infer anything of significance about the
infectivity of the virus, the Orf gene, which encodes the essential replicase enzyme of
SARS CoV viruses, should have been included as a target Figure . The positioning of the
targets in the region of the viral genome that is most heavily and variably transcribed is
another weakness of the protocol.
Kim et al. demonstrate a highly variable ’ expression of subgenomic RNA in Sars CoV .
These RNAs are actively monitored as signatures for asymptomatic and non infectious
patients . It is highly questionable to screen a population of asymptomatic people with
qPCR primers that have base pairs primer dimer on the prime end of a primer Figure .
Apparently the WHO recommends these primers. We tested all the wobble derivatives from
the Corman Drosten paper with Thermofisher’s primer dimer web tool . The RdRp
forward primer has bp prime homology with Sarbeco E Reverse. At high primer
concentrations this is enough to create inaccuracies.
Of note: There is a perfect match of one of the N primers to a clinical pathogen Pantoea ,
found in immuno compromised patients. The reverse primer hits Pantoea as well but not in
the same region Figure .
These are severe design errors, since the test cannot discriminate between the whole virus
and viral fragments. The test cannot be used as a diagnostic for SARS viruses.
Figure : Relative positions of amplicon targets on the SARS coronavirus and the novel coronavirus
genome. ORF: open reading frame; RdRp: RNA dependent RNA polymerase. Numbers below amplicon are
genome positions according to SARS CoV, NC ;
Figure : A test with Thermofischer’s primer dimer web tool reveals that the RdRp forward primer has a bp
`prime homology with Sarbeco E Reverse left box . Another test reveals that there is a perfect match for one
of the N primers to a clinical pathogen Pantoea found in immuno compromised patients right box .
. Reaction temperature
a DNA mel ing em e a e

Adequately addressed in the Corman Drosten paper.
b DNA am lifica ion em e a e

Adequately addressed in the Corman Drosten paper.
c E oneo GC con en and Tm

The annealing temperature determines at which temperature the primer attaches detaches
from the target sequence. For an efficient and specific amplification, GC content of primers
should meet a minimum of and a maximum of amplification. As indicated in table
, three of the primers described in the Corman Drosten paper are not within the normal
range for GC content. Two primers RdRp SARSr F and RdRp SARSr R have unusual and
very low GC values of for all possible variants of wobble bases, whereas primer
E Sarbeco F has a GC value of . Table and second panel of Table .
It should be noted that the GC content largely determines the binding to its specific target
due to its three hydrogen bonds in base pairing. Thus, the lower the GC content of the
primer, the lower its binding capability to its specific target gene sequence i.e. the gene to
be detected . This means for a target sequence to be recognized we have to choose a
temperature which is as close as possible to the actual annealing temperature best
practise value for the primer not to detach again, while at the same time specifically
selecting the target sequence.
If the Tm value is very low, as observed for all wobbly variants of the RdRp reverse primers,
the primers can bind non specifically to several targets, decreasing specificity and increasing
potential false positive results.
The annealing temperature Tm is a crucial factor for the determination of the

specificity accuracy of the qPCR procedure and essential for evaluating the accuracy of
qPCR protocols. Best practice recommendation: Both primers forward and reverse should
have an almost similar value, preferably the identical value.
We used the freely available primer design software Primer BLAST , to evaluable the
best practise values for all primers used in the Corman Drosten paper Table . We
attempted to find a Tm value of C, while similarly seeking the highest possible
GC value for all primers. A maximal Tm difference of C within primer pairs was
considered acceptable. Testing the primer pairs specified in the Corman Drosten paper, we
observed a difference of C with respect to the annealing temperature Tm for primer
pair RdRp SARSr F and RdRp SARSr R . This is a very serious error and makes the
protocol useless as a specific diagnostic tool.
Additional testing demonstrated that only the primer pair designed to amplify the N gene
N Sarbeco F and N Sarbeco R reached the adequate standard to operate in a diagnostic
test, since it has a sufficient GC content and the Tm difference between the primers
N Sarbeco F and N Sarbeco R is . C below the crucial maximum of C difference .
Importantly, this is the gene which was neither tested in the virus samples Table nor
emphasized as a confirmatory test. In addition to highly variable melting temperatures and
degenerate sequences in these primers, there is another factor impacting specificity of the
procedure: the dNTPs . uM are x higher than recommended for a highly specific
amplification. There is additional magnesium sulphate added to the reaction as well. This
procedure combined with a low annealing temperature can create non specific
amplifications. When additional magnesium is required for qPCR, specificity of the assay
should be further scrutinized.
The design errors described here are so severe that it is highly unlikely that specific
amplification of SARS CoV genetic material will occur using the protocol of the
Corman Drosten paper.
Table : GC content of the primers and probes adapted from Corman Drosten paper; aberrations from
optimized GC contents are highlighted. Second Panel shows a table listing of all Primer BLAST best practices
values for all primers and probes used in the Corman Drosten paper by Prof. Dr. Ulrike Kämmerer her team.
. The number of amplification cycles
It should be noted that there is no mention anywhere in the Corman Drosten paper of a test
being positive or negative, or indeed what defines a positive or negative result. These types
of virological diagnostic tests must be based on a SOP, including a validated and fixed
number of PCR cycles Ct value after which a sample is deemed positive or negative. The
maximum reasonably reliable Ct value is cycles. Above a Ct of cycles, rapidly increasing
numbers of false positives must be expected .
PCR data evaluated as positive after a Ct value of cycles are completely unreliable.
Citing Jaafar et al. :
A C he al e e ed o e o a o i i e e l fo PCR of c l e ae
oii e
In other words, there was no successful virus isolation of SARS CoV at those high Ct
values. Further, scientific studies show that only non infectious dead viruses are detected
with Ct values of .
Between and there is a grey area, where a positive test cannot be established with
certainty. This area should be excluded. Of course, one could perform PCR cycles, as
recommended in the Corman Drosten WHO protocol Figure , but then you also have to
define a reasonable Ct value which should not exceed . But an analytical result with a Ct
value of is scientifically and diagnostically absolutely meaningless a reasonable Ct value
should not exceed . All this should be communicated very clearly. It is a significant
mistake that the Corman Drosten paper does not mention the maximum Ct value at which
a sample can be unambiguously considered as a positive or a negative test result. This
important cycle threshold limit is also not specified in any follow up submissions to date.
Figure : RT PCR Kit recommendation in the official Corman Drosten WHO protocol . Only a Cycler value
cycles is to be found without corresponding and scientifically reasonable Ct Cutoff value . This or any other
cycles value is nowhere to be found in the actual Corman Drosten paper.
. Biomolecular validations
To determine whether the amplified products are indeed SARS CoV genes, biomolecular
validation of amplified PCR products is essential. For a diagnostic test, this validation is an
absolute must.
Validation of PCR products should be performed by either running the PCR product in a
agarose EtBr gel together with a size indicator DNA ruler or DNA ladder so that the size of
the product can be estimated. The size must correspond to the calculated size of the
amplification product. But it is even better to sequence the amplification product. The
latter will give certainty about the identity of the amplification product. Without
molecular validation one can not be sure about the identity of the amplified PCR products.
Considering the severe design errors described earlier, the amplified PCR products can be
anything.
Also not mentioned in the Corman Drosten paper is the case of small fragments of qPCR
around bp : It could be either , agarose gel or even an acrylamide gel.
The fact that these PCR products have not been validated at molecular level is another
striking error of the protocol, making any test based upon it useless as a specific diagnostic
tool to identify the SARS CoV virus.
. Positive and negative controls to confirm/refute specific virus detection.
The unconfirmed assumption described in the Corman Drosten paper is that SARS CoV is
the only virus from the SARS like beta coronavirus group that currently causes infections in
humans. The sequences on which their PCR method is based are in silico sequences,
supplied by a laboratory in China , because at the time of development of the PCR test
no control material of infectious live or inactivated SARS CoV was available to the
authors. The PCR test was therefore designed using the sequence of the known SARS CoV
as a control material for the Sarbeco component Dr. Meijer, co author Corman Drosten
paper in an email exchange with Dr. Peter Borger .
All individuals testing positive with the RT PCR test, as described in the Corman Drosten
paper, are assumed to be positive for SARS CoV infections. There are three severe flaws
in their assumption. First, a positive test for the RNA molecules described in the
Corman Drosten paper cannot be equated to infection with a virus . A positive RT PCR test
merely indicates the presence of viral RNA molecules. As demonstrated under point d
above , the Corman Drosten test was not designed to detect the full length virus, but only
a fragment of the virus. We already concluded that this classifies the test as unsuitable as a
diagnostic test for SARS virus infections.
Secondly and of major relevance, the functionality of the published RT PCR Test was not
demonstrated with the use of a positive control isolated SARS CoV RNA which is an
essential scientific gold standard.
Third, the Corman Drosten paper states:
To ho ha he a a can de ec o he ba a ocia ed SARS ela ed i e e ed he E

gene a a o e i ba de i ed faecal am le a ailable f om D e le e al … nd M h
e al … The e i o i i e am le emmed f om E o ean hinolo hid ba De ec ion
of he e h logene ic o lie i hin he SARS ela ed CoV clade gge ha all A ian
i e a e likel o be de ec ed Thi o ld heo e icall en e b oad en i i i e en in
ca e of m l i le inde enden ac i i ion of a ian i e f om an animal e e oi
This statement demonstrates that the E gene used in RT PCR test, as described in the
Corman Drosten paper, is not specific to SARS CoV .
The E gene primers also detect a broad spectrum of other SARS viruses.
The genome of the coronavirus is the largest of all RNA viruses that infect humans and they
all have a very similar molecular structure. Still, SARS CoV and SARS CoV have two highly
specific genetic fingerprints, which set them apart from the other coronaviruses. First, a
unique fingerprint sequence KTFPPTEPKKDKKKK is present in the N protein of SARS CoV
and SARS CoV , , . Second, both SARS CoV and SARS CoV do not contain the HE
protein, whereas all other coronaviruses possess this gene , . So, in order to
specifically detect a SARS CoV and SARS CoV PCR product the above region in the N gene
should have been chosen as the amplification target. A reliable diagnostic test should focus
on this specific region in the N gene as a confirmatory test. The PCR for this N gene was not
further validated nor recommended as a test gene by the Drosten Corman paper, because of
being not so sensitive with the SARS CoV original probe .
Furthermore, the absence of the HE gene in both SARS CoV and SARS CoV makes this
gene the ideal negative control to exclude other coronaviruses. The Corman Drosten paper
does not contain this negative control, nor does it contain any other negative controls. The
PCR test in the Corman Drosten paper therefore contains neither a unique positive control
nor a negative control to exclude the presence of other coronaviruses. This is another major
design flaw which classifies the test as unsuitable for diagnosis.
. Standard Operational Procedure SOP is not available
There should be a Standard Operational Procedure SOP available, which unequivocally

specifies the above parameters, so that all laboratories are able to set up the identical same
test conditions. To have a validated universal SOP is essential, because it facilitates data
comparison within and between countries. It is very important to specify all primer
parameters unequivocally. We note that this has not been done. Further, the Ct value to
indicate when a sample should be considered positive or negative is not specified. It is also
not specified when a sample is considered infected with SARS CoV viruses. As shown above,
the test cannot discern between virus and virus fragments, so the Ct value indicating
positivity is crucially important. This Ct value should have been specified in the Standard
Operational Procedure SOP and put on line so that all laboratories carrying out this test
have exactly the same boundary conditions. It points to flawed science that such an SOP
does not exist. The laboratories are thus free to conduct the test as they consider
appropriate, resulting in an enormous amount of variation. Laboratories all over Europe are
left with a multitude of questions; which primers to order? which nucleotides to fill in the
undefined places? which Tm value to choose? How many PCR cycles to run? At what Ct value
is the sample positive? And when is it negative? And how many genes to test? Should all
genes be tested, or just the E and RpRd gene as shown in Table of the Corman Drosten
paper? Should the N gene be tested as well? And what is their negative control? What is
their positive control?
The protocol as described is unfortunately very vague and erroneous in its design that one
can go in dozens of different directions. There does not appear to be any standardization nor
an SOP, so it is not clear how this test can be implemented.
. Consequences of the errors described under - : false positive results.
The RT PCR test described in the Corman Drosten paper contains so many molecular
biological design errors see that it is not possible to obtain unambiguous results. It is
inevitable that this test will generate a tremendous number of so called false positives .
The definition of false positives is a negative sample, which initially scores positive, but
which is negative after retesting with the same test. False positives are erroneous positive
test results, i.e. negative samples that test positive. And this is indeed what is found in the
Corman Drosten paper. On page of the manuscript PDF the authors demonstrate, that
even under well controlled laboratory conditions, a considerable percentage of false
positives is generated with this test:
In fo indi id al e eac ion eak ini ial eac i i a een ho e e he e e nega i e

on e e ing i h he ame a a The e ignal e e no a ocia ed i h an a ic la
i and fo each i i h hich ini ial o i i e eac i i occ ed he e e e o he
am le ha con ained he ame i a a highe concen a ion b did no e o i i e
Gi en he e l f om he e en i e echnical alifica ion de c ibed abo e i a concl ded
ha hi ini ial eac i i a no d e o chemical in abili of eal ime PCR obe and
mo obabl o handling i e ca ed b he a id in od c ion of ne diagno ic e
and con ol d ing hi e al a ion d
The first sentence of this excerpt is clear evidence that the PCR test described in the
Corman Drosten paper generates false positives. Even under the well controlled conditions
of the state of the art Charité laboratory, out of primary tests are false positives per
definition. Four negative samples initially tested positive, then were negative upon retesting.
This is the classical example of a false positive. In this case the authors do not identify them
as false positives, which is intellectually dishonest.
Another telltale observation in the excerpt above is that the authors explain the false
positives away as handling issues caused by the rapid introduction of new diagnostic tests .
Imagine the laboratories that have to introduce the test without all the necessary
information normally described in an SOP.
. The Corman-Drosten paper was not peer-reviewed
Before formal publication in a scholarly journal, scientific and medical articles are
traditionally certified by peer review. In this process, the journal’s editors take advice from
various experts referees who have assessed the paper and may identify weaknesses in its
assumptions, methods, and conclusions. Typically a journal will only publish an article once
the editors are satisfied that the authors have addressed referees’ concerns and that the
data presented supports the conclusions drawn in the paper. This process is as well
described for Eurosurveillance .
The Corman Drosten paper was submitted to Eurosurveillance on January st and

accepted for publication on January nd . On January rd the paper was online.
On January th version of the protocol was published at the official WHO website
, updated on January th as document version , even before the
Corman Drosten paper was published on January rd at Eurosurveillance.
Normally, peer review is a time consuming process since at least two experts from the field
have to critically read and comment on the submitted paper. In our opinion, this paper was
not peer reviewed. Twenty four hours are simply not enough to carry out a thorough peer
review. Our conclusion is supported by the fact that a tremendous number of very serious
design flaws were found by us, which make the PCR test completely unsuitable as a
diagnostic tool to identify the SARS CoV virus. Any molecular biologist familiar with RT PCR
design would have easily observed the grave errors present in the Corman Drosten paper
before the actual review process. We asked Eurosurveillance on October th to send
us a copy of the peer review report. To date, we have not received this report and in a letter
dated November th , the ECDC as host for Eurosurveillance declined to provide
access without providing substantial scientific reasons for their decision. On the contrary,
they write that disclosure would undermine the purpose of scientific investigations. .
. Authors as the editors
A final point is one of major concern. It turns out that two authors of the Corman Drosten
paper, Christian Drosten and Chantal Reusken, are also members of the editorial board of
this journal . Hence there is a severe conflict of interest which strengthens suspicions
that the paper was not peer reviewed. It has the appearance that the rapid publication was
possible simply because the authors were also part of the editorial board at
Eurosurveillance. This practice is categorized as compromising scientific integrity.
SUMMARY CATALOGUE OF ERRORS FOUND IN THE PAPER
The Corman Drosten paper contains the following specific errors:
. There exists no specified reason to use these extremely high concentrations of primers in
this protocol. The described concentrations lead to increased nonspecific bindings and PCR
product amplifications, making the test unsuitable as a specific diagnostic tool to identify the
SARS CoV virus.
. Six unspecified wobbly positions will introduce an enormous variability in the real world
laboratory implementations of this test; the confusing nonspecific description in the
Corman Drosten paper is not suitable as a Standard Operational Protocol making the test
unsuitable as a specific diagnostic tool to identify the SARS CoV virus.
. The test cannot discriminate between the whole virus and viral fragments. Therefore, the
test cannot be used as a diagnostic for intact infectious viruses, making the test unsuitable
as a specific diagnostic tool to identify the SARS CoV virus and make inferences about the
presence of an infection.
. A difference of C with respect to the annealing temperature Tm for primer pair

RdRp SARSr F and RdRp SARSr R also makes the test unsuitable as a specific diagnostic
. A severe error is the omission of a Ct value at which a sample is considered positive and
negative. This Ct value is also not found in follow up submissions making the test unsuitable
as a specific diagnostic tool to identify the SARS CoV virus.
. The PCR products have not been validated at the molecular level. This fact makes the
protocol useless as a specific diagnostic tool to identify the SARS CoV virus.
. The PCR test contains neither a unique positive control to evaluate its specificity for
SARS CoV nor a negative control to exclude the presence of other coronaviruses, making
the test unsuitable as a specific diagnostic tool to identify the SARS CoV virus.
. The test design in the Corman Drosten paper is so vague and flawed that one can go in
dozens of different directions; nothing is standardized and there is no SOP. This highly
questions the scientific validity of the test and makes it unsuitable as a specific diagnostic
. Most likely, the Corman Drosten paper was not peer reviewed making the test
unsuitable as a specific diagnostic tool to identify the SARS CoV virus.
. We find severe conflicts of interest for at least four authors, in addition to the fact that
two of the authors of the Corman Drosten paper Christian Drosten and Chantal Reusken
are members of the editorial board of Eurosurveillance. A conflict of interest was added on
July Olfert Landt is CEO of TIB Molbiol; Marco Kaiser is senior researcher at
GenExpress and serves as scientific advisor for TIB Molbiol , that was not declared in the
original version and still is missing in the PubMed version ; TIB Molbiol is the company
which was the first to produce PCR kits Light Mix based on the protocol published in the
Corman Drosten manuscript, and according to their own words, they distributed these
PCR test kits before the publication was even submitted ; further, Victor Corman
Christian Drosten failed to mention their second affiliation: the commercial test laboratory
Labor Berlin . Both are responsible for the virus diagnostics there and the company
operates in the realm of real time PCR testing.
In light of our re-examination of the test protocol to identify SARS-CoV- described in the
Corman-Drosten paper we have identified concerning errors and inherent fallacies which
render the SARS-CoV- PCR test useless.
CONCLUSION
The decision as to which test protocols are published and made widely available lies
squarely in the hands of Eurosurveillance. A decision to recognise the errors apparent in the
Corman Drosten paper has the benefit to greatly minimise human cost and suffering going
forward.
Is it not in the best interest of Eurosurveillance to retract this paper? Our conclusion is
clear. In the face of all the tremendous PCR protocol design flaws and errors described
here, we conclude: There is not much of a choice left in the framework of scientific integrity
and responsibility.
REFERENCES
Corman Victor M, Landt Olfert, Kaiser Marco, Molenkamp Richard, Meijer Adam, Chu Daniel KW, Bleicker
Tobias, Brünink Sebastian, Schneider Julia, Schmidt Marie Luisa, Mulders Daphne GJC, Haagmans Bart L, van
der Veer Bas, van den Brink Sharon, Wijsman Lisa, Goderski Gabriel, Romette Jean Louis, Ellis Joanna, Zambon
Maria, Peiris Malik, Goossens Herman, Reusken Chantal, Koopmans Marion PG, Drosten Christian. Detection
of novel coronavirus nCoV by real time RT PCR. Euro Surveill. ; :pii .
https: doi.org . .ES. . . .
Email communication between Dr. Peter Borger Dr. Adam Meijer: Supplementary Material
Jafaar et al., Correlation Between Quantitative Polymerase Chain Reaction Positives Samples and
Positive Cell Cultures, Including Severe Acute Respiratory Syndrome Coronavirus Isolates.
https: academic.oup.com cid advance article doi . cid ciaa
BBC, January st : https: www.bbc.com news world asia china ;
Archive: https: archive.is qRmZ
Google Analytics COVID deaths worldwide: https: bit.ly fndemJ
Archive: https: archive.is PpqEE
Laboratory testing for COVID Emergency Response Technical Centre, NIVD under
China CDC March th, : http: www.chinacdc.cn en COVID P .pdf
Real Time PCR Handbook Life Technologies:

https: www.thermofisher.com content dam LifeTech global Forms PDF real time pcr
handbook.pdf
Nolan T, Huggett J, Sanchez E.Good practice guide for the application of quantitative PCR qPCR First Edition
Trestan Pillonel et al, Letter to the editor: SARS CoV detection by real time RT PCR:
https: www.ncbi.nlm.nih.gov pmc articles PMC
Kurkela, Satu, and David WG Brown. Molecular diagnostic techniques. Medicine . : .
Wolfel et al., Virological assessment of hospitalized patients with COVID
https: www.nature.com articles s x
Thermofischer Primer Dimer Web Tool:

https: www.thermofisher.com us en home brands thermo scientific molecular biology molecular biology
learning center molecular biology resource library thermo scientific web tools multiple primer analyzer.h
tml
Suplementary material by Kevin Mckernan, Corman Drosten Primer Dimer Results with Thermofischer
Primer Dimer Web Tool
Primer BLAST, NCBI National Center for Biotechnology Information:

https: www.ncbi.nlm.nih.gov tools primer blast
Marra MA, Steven JMJ, Caroline RA, Robert AH, Angela BW et al. Science. The
Genome sequence of the SARS associated coronavirus. Science : .
Severe acute respiratory syndrome coronavirus isolate Wuhan Hu , complete genome:

https: www.ncbi.nlm.nih.gov nuccore MN
Borger P. A SARS like Coronavirus was expected but nothing was done to be prepared. Am J Biomed Sci
Res . https: biomedgrid.com pdf AJBSR.MS.ID. .pdf
https: www.researchgate.net publication A SARS like Coronavirus was Expected but nothi
ng was done to be Prepared;
Archive: https: archive.is i Hu
Eurosurveillance paper evaluation review process: https: www.eurosurveillance.org evaluation
Official recommendation of the Corman Drosten protocol manuscript by the WHO,published on

January th as version . of the document:
https: www.who.int docs default source coronaviruse wuhan virus assay v e d a b

c ttps: bit.ly m jXVH
.pdf; archive: h
Official WHO recommendation for the Corman Drosten RT qPCR protocol, which directly derives from
the Eurosurveillance publication, document version , published on
th January :
https: www.who.int docs default source coronaviruse protocol v .pdf?sfvrsn a ef c
Eurosurveillance Editorial Board, :

https: www.eurosurveillance.org upload site assets imgs Editorial Board PDF.pdf;
Archive: https: bit.ly TqXBjX
Instructions For Use LightMix SarbecoV E gene plus EAV Control, TIB Molbiol Roche Molecular
Solutions, January th :
https: www.roche as.es lm pdf MDx Sarbeco E gene V .pdf
Archive, timestamp January th : https: archive.is Vulo ;
Archive: https: bit.ly fm bXH
Christian Drosten Victor Corman, responsible for viral diagnostics at Labor Berlin:
https: www.laborberlin.com fachbereiche virologie
Archive: https: archive.is CDEUG
Tom Jefferson, Elizabeth Spencer, Jon Brassey, Carl Heneghan Viral cultures for COVID infectivity
assessment. Systematic review. Systematic review doi: https: doi.org . . . .
https: www.medrxiv.org content . . . . v
Kim et al.,The Architecture of SARS CoV Transcriptome:

https: www.sciencedirect.com science article pii S
ECDC reply to Dr. Peter Borger, th November : Supplementary Material
Prof. Dr. Ulrike Kämmerer team, survey Primer BLAST table: Supplementary Material
Additional literature:
Description RT PCR RKI Germany, on page of this link:

https: www.rki.de DE Content Gesundheitsmonitoring Gesundheitsberichterstattung GBEDownloadsJ Jo
HM S Studienprotokoll CORONA MONITORING lokal.pdf? blob publicationFile
Author s Affiliations:
Dr. Pieter Borger MSc, PhD , Molecular Genetics, W W Research Associate, Lörrach, Germany
Rajesh Kumar Malhotra Artist Alias: Bobby Rajesh Malhotra , Former D Artist / Scientific Visualizations at CeMM - Center for Molecular Medicine of the
Austrian Academy of Sciences - , University for Applied Arts - Department for Digital Arts Vienna, Austria
Dr. Michael Yeadon BSs Hons Biochem Tox U Surrey, PhD Pharmacology U Surrey. Managing Director, Yeadon Consulting Ltd, former Pfizer Chief
Scientist, United Kingdom
Dr. Clare Craig MA, Cantab BM, BCh Oxon , FRCPath, United Kingdom
Kevin McKernan, BS Emory University, Chief Scientific Officer, founder Medical Genomics, engineered the sequencing pipeline at WIBR/MIT for the
Human Genome Project, Invented and developed the SOLiD sequencer, awarded patents related to PCR, DNA Isolation and Sequencing, USA
Prof. Dr. Klaus Steger, Department of Urology, Pediatric Urology and Andrology, Molecular Andrology, Biomedical Research Center of the Justus Liebig
University, Giessen, Germany
Dr. Paul McSheehy BSc, PhD , Biochemist Industry Pharmacologist, Loerrach, Germany
Dr. Lidiya Angelova, MSc in Biology, PhD in Microbiology, Former researcher at the National Institute of Allergy and Infectious Diseases NIAID ,
Maryland, USA
Dr. Fabio Franchi, Former Dirigente Medico M.D in an Infectious Disease Ward, specialized in “Infectious Diseases” and “Hygiene and Preventive
Medicine”, Società Scientifica per il Principio di Precauzione SSPP , Italy
Dr. med. Thomas Binder, Internist and Cardiologist FMH , Switzerland
Prof. Dr. med. Henrik Ullrich, specialist Diagnostic Radiology, Chief Medical Doctor at the Center for Radiology of Collm Oschatz-Hospital, Germany
Prof. Dr. Makoto Ohashi, Professor emeritus, PhD in Microbiology and Immunology, Tokushima University, Japan
Dr. Stefano Scoglio, B.Sc. Ph.D., Microbiologist, Nutritionist, Italy
Dr. Marjolein Doesburg-van Kleffens MSc, PhD , specialist in Laboratory Medicine clinical chemistry , Maasziekenhuis Pantein, Beugen, The
Netherlands
Dr. Dorothea Gilbert MSc, PhD , PhD Environmental Chemistry and Toxicology. DGI Consulting Services, Oslo, Norway
Dr. Rainer J. Klement, PhD. Department of Radiation Oncology, Leopoldina Hospital Schweinfurt, Germany
Dr. Ruth Schruefer, PhD, human genetics/ immunology, Munich, Germany,
Dra. Berber W. Pieksma, General Practitioner, The Netherlands
Dr. med. Jan Bonte GJ , Consultant Neurologist, The Netherlands
Dr. Bruno H. Dalle Carbonare Molecular biologist , IP specialist, BDC Basel, Switzerland
Dr. Kevin P. Corbett, MSc Nursing Kings College London PhD London South Bank Social Sciences Science Technology Studies London, England,
United Kingdom
Prof. Dr. Ulrike Kämmerer, specialist in Virology / Immunology / Human Biology / Cell Biology, University Hospital Würzburg, Germany
Author’s Contributions:
PB: Planned and conducted the analyses and research, conceptualising the manuscript.
BRM: Planned and conducted the research, conceptualising the figures and manuscript.
MY: Conducted the analyses and research.
KMcK: Conducted the analyses and research, conceptualized the manuscript.
KS: Conducted the analyses and research.
PMcS: Proofreading the analyses and research.
LA: Proofreading the analyses and research.
FF: Proofreading the analyses and research.
TB: Proofreading the analyses and research.
HU: Proofreading the analyses and research.
MO: Proofreading the analyses and research.
SS: Proofreading the analyses and research.
MDvK: Proofreading the analyses and research.
DG: Proofreading the analyses and research.
RJK: Proofreading the analyses and research.
RS: Proofreading the analyses and research, and the manuscript.
BWK: Proofreading the analyses and research.
RvV: Proofreading the analyses and research.
JB: Proofreading the analyses and research.
KC: Proofreading the analyses and research.
UK: Planned and conducted the analyses and research, conceptualising the manuscript.
Additional Proof-Readers:
Saji N Hameed, Environmental Informatics, University of Aizu, Tsuruga, Ikki-machi, Aizuwakamatsu-shi, Fukushima, Japan
Howard R. Steen, MA Chem. Eng. Cantab, Former Research Manager, Germany


Wodarg Yeadon EMA Petition Pfizer Trial FINAL 01DEC2020 en Unsigned With Exhibits

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Wodarg Yeadon EMA Petition Pfizer Trial FINAL 01DEC2020 en Unsigned With Exhibits

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PETITIONER: December 1, 2020

Dr. med. Wolfgang Wodarg

England, CT3 1RT

PHASE III - EUDRACT NUMBER: 2020-002641-42

SPONSOR PROTOCOL NUMBER: C4591001

AND ANY OTHER ONGOING CLINICAL TRIALS OF VACCINE CANDIDATES

ADMINISTRATIVE/REGULATORY STAY OF ACTION

Before an Emergency Authorization/Conditional Approval and/or Unrestricted

Before an Emergency Authorization/Conditional Approval and/or Unrestricted

X. In the Pfizer/BioNTech mRNA vaccine candidate, polyethylene glycol (PEG) is found in

According to section 10.4.2 of the Pfizer/BioNTech trial protocol, a woman of childbearing

Respectfully submitted on my behalf and on behalf of Co-Petitioner Dr. Michael Yeadon:

November 25, 2020

Division of Dockets Management

UNITED STATES DEPARTMENT OF HEALTH AND HUMAN SERVICES

PETITION FOR ADMINISTRATIVE :

ADMINISTRATIVE STAY OF ACTION

1. Approval of trial design for Phase III trial of BNT162 (NCT04368728)1

Before an EUA or unrestricted license is issued for the Pfizer

All RT-qPCR-positive test results used to categorize patient as

7. Furthermore, if the vaccine is approved without an appropriate and accurate review

7. According to the trial protocol, 8.1. Efficacy and/or Immunogenicity

presence of at least 1 of the following symptoms and SARS-CoV-2 NAAT-positive

10. According to the trial protocol, 8.1 Efficacy and/or Immunogenicity

Cepheid Xpert Xpress SARS-CoV-2

h. DNA sequencing verification is necessary for confirmation of the presumptive SARS-

5400 Cepheid Xpert Xpress SARS-CoV-2 test

5400 Abbott Molecular Abbott RealTime SARS-CoV-2 assay

Fever 39.0°C ( 102.1°F).

Stay Urgently Required

Dr. Sin Hang Lee

External peer review of the RTPCR test to detect SARS-CoV-

CONCISE REVIEW REPORT

We aimed o de elo and de lo ob diagno ic me hodolog fo e in blic

he e abli hmen and alida ion of a diagno ic o kflo fo nCoV c eening

The Reverse Transcription Polymerase Chain Reaction RT PCR is an important biomolecular

. The primers and probes:

a the concentration of primers and probes must be of optimal range nM

b must be specific to the target gene you want to amplify

. The temperature at which all reactions take place:

a DNA melting temperature

b DNA amplification temperature TaqPol specific

. The number of amplification cycles less than ; preferably - cycles ;

. There should be a Standard Operational Procedure SOP available

MINOR CONCERNS WITH THE CORMAN-DROSTEN PAPER

MAJOR CONCERNS WITH THE CORMAN-DROSTEN PAPER

B METHODS AND RESULTS

. Primer Probe Design

a Erroneous primer concentrations

b Unspecified “Wobbly primer and probe sequences

either an A or a T; R signifies there can be either a G or an A; M indicates that the position

Of no e he N gene a a al o e fo med ell b a no bjec ed

c Erroneous GC content discussed in c, together with annealing temperature Tm

d Detection of viral genes

Clinician need o ecogni e he enhanced acc ac and eed of he molec la diagno ic

a DNA mel ing em e a e

b DNA am lifica ion em e a e

c E oneo GC con en and Tm

The annealing temperature Tm is a crucial factor for the determination of the

. The number of amplification cycles

Citing Jaafar et al. :

. Positive and negative controls to confirm/refute specific virus detection.

Third, the Corman Drosten paper states:

To ho ha he a a can de ec o he ba a ocia ed SARS ela ed i e e ed he E

BBC, January st : https: www.bbc.com news world asia china ;

Archive: https: archive.is qRmZ

Google Analytics COVID deaths worldwide: https: bit.ly fndemJ

Archive: https: archive.is PpqEE

China CDC March th, : http: www.chinacdc.cn en COVID P .pdf

Archive: https: archive.is i Hu

Eurosurveillance paper evaluation review process: https: www.eurosurveillance.org evaluation

Archive: https: bit.ly TqXBjX

Archive, timestamp January th : https: archive.is Vulo ;

Archive: https: bit.ly fm bXH

Archive: https: archive.is CDEUG

ECDC reply to Dr. Peter Borger, th November : Supplementary Material