Molecular detection of novel coronaviruses

Abstract

Systems for sensitive and accurate detection of sarbecoviruses in samples from human sputum have been established. The systems can rapidly and effectively diagnose patients having suspected coronavirus infections. The systems employ Reverse-Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) to identify and quantify nucleic acids specific to sarbecoviruses within samples from human patients. The systems identify viral RNAs specific for the N and ORF1 genes using a two-step real-time PCR and the SYBR Green detection method. Compositions and methods for reproducibly detecting and quantifying concentrations of sarbecoviruses with a dynamic range of at least seven orders of magnitude (2×10−4-2000 TCID50/reaction) are described. The compositions and methods enable rapid early diagnosis of patients, as well as development of databases of patient viral titres and infectious dosages.

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