RT-qPCR Diagnostics: The “Drosten” SARS-CoV-2 Assay Paradigm

The reverse transcription quantitative polymerase chain reaction (RT-qPCR) is an established tool for the diagnosis of RNA pathogens. Its potential automation has caused it to be used as a presence/absence diagnostic even when quantification not required. This technology been pushed forefront public awareness by COVID-19 pandemic, its global application enabled rapid and analytically sensitive mass testing, with first assays targeting three viral genes published within days publication SARS-CoV-2 genomic sequence. One those, RNA-dependent gene, heavily criticised supposed scientific flaws at molecular methodological level, this criticism extrapolated doubts about validity RT-qPCR testing in general. We have analysed assay detail, our findings reveal some limitations but also highlight robustness methodology detection. Nevertheless, whilst data show that errors can tolerated, always prudent confirm primer probe sequences complement their intended target, since, do occur, they may result reduction analytical sensitivity. However, case, unlikely mismatch will poor specificity or significant number false-positive diagnoses, especially routinely checked laboratories part quality assurance.