"Abstract: In the publication entitled 'Detection of 2019 novel coronavirus(2019-nCoV) by real-time RT-PCR' (Eurosurveillance 25(8) 2020), the authors present a diagnostic workflow and RT-qPCR protocol for detection and diagnostics of 2019-nCoV (now known as SARS-CoV-2), 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 1) all components of the presented test design were cross checked, 2) the RT-qPCR protocol-recommendations were assessed w.r.t. good laboratory practice, and 3) parameters examined against relevant scientific literature covering the field.
The published RT-qPCR protocol for detection and diagnostics of 2019-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 fulfills the requirements for an acceptable scientific publication...
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.
What is important when designing an RT-PCR Test and the quantitative RT-qPCR test described in the Corman-Drosten publication?…
Concise Review Report
... There are ten fatal problems with the Corman-Drosten paper which we will outline and explain in greater detail in the following sections…
3. The number of amplification cycles (less than 35; preferably 25-30 cycles); In case of virus detection, >35 cycles only detects signals which do not correlate with infectious virus as determined by isolation in cell culture [reviewed in 2]; if someone is tested by PCR as positive when a threshold of 35 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 3%, the probability that said result is a false positive is 97%…
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…
Further, scientific studies show that only non-infectious (dead) viruses are detected with Ct values of 35.”
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