De Corman-Drosten Peer Reviewed studie over PCR-testen ingediend bij Eurosurveillance
Velen onder jullie hebben mij berichten gestuurd met meldingen van alternatieve media kanalen over de " retraction paper " inzake PCR testen, dat ik NIET gepost hebben omdat het slechts mededelingen waren.
Hier vindt u de link naar de retraction letter zelve : https://cormandrostenreview.com/retraction-request.../
En hier vindt u de link naar de peer review https://cormandrostenreview.com/report/...
Vaststellingen :
1. Primer & Probe Design 1a) Erroneous primer concentrations
1b) Unspecified (“Wobbly”) primer and probe sequences
1c) Erroneous GC-content (discussed in 2c, together with annealing temperature (Tm))
1d) 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-19 is not indicated for COVID-19 diagnosis on a regulatory basis.
2. Reaction temperatures 2a) DNA melting temperature (>92°). Adequately addressed in the Corman-Drosten paper. 2b) DNA amplification temperature. Adequately addressed in the Corman-Drosten paper. 2c) Erroneous GC-contents and Tm
3. 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 30 cycles. Above a Ct of 35 cycles, rapidly increasing numbers of false positives must be expected .
PCR data evaluated as positive after a Ct value of 35 cycles are completely unreliable.
4. Biomolecular validations 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-2 virus.
5. Positive and negative controls to confirm/refute specific virus detection. 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. the E gene used in RT-PCR test, as described in the Corman-Drosten paper, is not specific to SARS-CoV-2. in order to specifically detect a SARS-CoV1 and SARS-CoV-2 PCR product the above region in the N gene should have been chosen as the amplification target.
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.
6. Standard Operational Procedure (SOP) is not available 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.
7. Consequences of the errors described under 1-5: false positive results.
8. The Corman-Drosten paper was not peer-reviewed ( nu wel dus )
9. 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 [19]. Hence there is a severe conflict of interest which strengthens suspicions that the paper was not peer-reviewed.
The Corman-Drosten paper contains the following specific errors:
1. 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-2 virus.
2. 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-2 virus.
3. 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-2 virus and make inferences about the presence of an infection.
4. A difference of 10° C with respect to the annealing temperature Tm for primer pair1 (RdRp_SARSr_F and RdRp_SARSr_R) also makes the test unsuitable as a specific diagnostic tool to identify the SARS-CoV-2 virus.
5. 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-2 virus.
6. 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-2 virus.
7. The PCR test contains neither a unique positive control to evaluate its specificity for SARS-CoV-2 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-2 virus.
8. 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 tool to identify the SARS-CoV-2 virus.
9. Most likely, the Corman-Drosten paper was not peer-reviewed making the test unsuitable as a specific diagnostic tool to identify the SARS-CoV-2 virus.
10. 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 29 2020 (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 [20]; 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 [21] 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-2 described in the Corman-Drosten paper we have identified concerning errors and inherent fallacies which render the SARS-CoV-2 PCR test useless.
Bij samenlezing van deze peer review en de andere wetenschappellijke studies over PCR testen ( waaronder deze van het CDC - zie eerdere posts ) kan niet anders dan geconcludeerd worden dan dat de PCR test waardeloos is.
