METHOD FOR THE DETECTION OF SARS-COV-2 BY DIGITAL PCR

The comparison of the diagnostic sensitivity between RT-qPCR and RT-ddPCR for SARS-CoV-2 detection has revealed that implementation of RT-ddPCR eliminates false negative results and enables accurate and sensitive quantification of low viral load of SARS-CoV-2 when compared to RT-qPCR. Analytical and clinical validation of highly sensitive molecular assays is highly important for the reliable detection and quantification of SARS-COV-2 RNA transcripts, leading to better management of infected patients, confirmatory analysis of samples with high Cq values, and viral detection in wastewater samples. The present invention provides a highly sensitive and specific in vitro method for the quantification of SARS-CoV-2 in a sample comprising a one-step multiplex reverse transcription digital PCR (RT-dPCR) assay. The method of the present invention comprises a one-step multiplex RT-dPCR assay for detecting SARS-CoV-2 in a sample and involves the quantification of three different regions of the nucleoprotein (N) gene of SARS-CoV-2, the use of a non-natural synthetic RNA as RNA exogenous control and the use of an endogenous RNA internal control. The present invention further provides a kit for the quantification of SARS-CoV-2 in a sample.