Rapid Genotyping of SARS-CoV-2 Strains Alpha, Delta, Omicron BA.1, Omicron BA.2, Omicron BA.2.12.1, and Omicron BA.4/BA.5 Using ARMS- PCR and Molecular Beacon Technology (preprint)

ABSTRACT

Background SARS-CoV-2 subtypes Alpha, Delta, Omicron BA.1, Omicron BA.2, Omicron BA.2.12.1, and Omicron BA.4/BA.5 have significant differences in transmission and immune escape ability. Currently, no effective detection methods are available for these subtypes. Routine detection methods are prone to missed detection.Methods In this study, a rapid detection method based on ARMS-PCR and molecular beacon probes was developed for the identification of SARS-CoV-2 subtypes Alpha, Delta, Omicron BA.1, Omicron BA.2, Omicron BA.2.12.1, and Omicron BA.4/5. Specific primers and probes were designed and validated using gel electrophoresis, real-time fluorescence quantitative PCR, and molecular hybridization.Results ARMS-PCR and molecular beacon probe-based assays can be applied in RT-PCR and fluorescence assays to differentiate SARS-CoV-2 subtypes Alpha, Delta, Omicron BA.1, Omicron BA.2, Omicron BA.2.12.1, and Omicron BA.4/5.Conclusions In the present study, we developed a simple, rapid and accurate detection method based on ARMS-PCR and molecular beacon probes for rapid genotyping of SARS-CoV-2 subtypes Alpha, Delta, Omicron BA.1, Omicron BA.2, Omicron BA.2.12.1, and Omicron BA.4/5. It can be used in real-time fluorescence quantitative PCR and molecular hybridization to identify subtypes of COVID-19, effectively improving the detection rate to provide guidance for disease prevention and treatment.