CALL FOR ACCOUNTABILITY AND TRANSPARENCY: Outcomes of Pediatric SARS-CoV-2 Omicron Infection vs Influenza and Respiratory Syncytial Virus Infections (JAMA) Authors Should Publish FULL PCR Protocol
DR. LEE OF THE IPAK NAATEC CONSORTIUM PUBLISHES LETTER CALLING FOR PUBLICATION OF FULL PCR PROTOCOL ON PEDIATRIC RESPIRATORY VIRUS STUDY
[SOURCE]
December 28, 2023
Sin Hang Lee, MD | Milford Molecular Diagnostics Laboratory
Dr. Pontus Hedberg and colleagues used multiplex PCR to diagnose SARS-CoV-2, influenza A/B, or RSV infection among individuals younger than 18 years for a retrospective comparative study and suggested that RSV infections more often require hospitalization and respiratory support, underscoring the importance of preventive measures, such as recently approved RSV vaccines. Since multiplex PCR is known to generate false-negative and false-positive results [1] and a Sanger sequence containing a minimum of 100 contiguous bases matching the reference with an E-Value < 10(-30) for a BLAST search in GenBank is required for NAAT-based diagnosis of viral infections [2], the authors should publish their full PCR protocol, including the multiplex PCR agarose gel electrophoresis and the sequencing electropherograms to prove that their multiplex PCR products were in fact amplicons of the SARS-CoV-2, influenza A/B, or RSV genomic nucleic acid sequences to support their suggestion.
References
1. Henegariu O, Heerema NA, Dlouhy SR, Vance GH, Vogt PH. Multiplex PCR: critical parameters and step-by-step protocol. Biotechniques. 1997;23:504-11.
2. Food and Drug Administration. Establishing the Performance Characteristics of In Vitro Diagnostic Devices for the Detection or Detection and Differentiation of Human Papillomaviruses https://www.fda.gov/media/92930/download
Sin Hang Lee, MD
Shlee01 [at] snet.net
Member of the NAATEC Consortium
CONFLICT OF INTEREST: SHL is the director of Milford Molecular Diagnostics Laboratory, a CLIA-certified laboratory specializing in Sanger sequencing diagnostic technologies.
Dr Jack, you’re the perfect person to ask. I have a couple questions, if you’re willing, on pcr. I sent you a free book, so please forgive me if they are silly/dumb questions from a nurse. I tend to lean toward Mullis’ own words that it is not for clinical diagnostics so doubt usefulness, period. But, 1. what methods do you want them to publish that would specifically confirm, or certainly call into question, their findings? (I am trying to educate myself) 2. is anything beyond 16-20 or so cycles total hogwash regardless of setting or field of study? I ask because new labs are making bank, and (I wonder) fraudulently?, on pcr for all kinds of testing now.. swabs for wounds, urine, etc. I’ve long suspected. For example, a ‘clinical’ lab in widespread use now runs their urine for ‘culture’ at 40 cycles.
Her conflict of interest tells us she works in the field and makes me think she knows her stuff.
Many of the primers for PCR and lateral flow come from labs in China with management beholden to who knows where. Even if the labs were to follow GOOD practice the results could be rubbish if the reagents were compromised. I would like to see baseline/null/challenge results for all the reagents to see if they are sensitive to other random stuff even at low threasholds.