FIELD: medical microbiology; molecular biology.
SUBSTANCE: what is described is a method for detection and quantification of SARS-CoV-2 virus by means of a droplet digital PCR providing: recovery of total RNA, obtaining complementary DNA (cDNA) on its basis, conducting ddPCR using complementary to the stable fragment of SARS-CoV-2 gene S region primers and a probe SEQ ID NO: 1-3: SEQ ID NO: 1 PR 55 F – 5’-TTGCCACCTTTGCTCACAGA-3’, SEQ ID NO: 2 PR 56 R – 5’-AAAGGTCCAACCAGAAGTGA-3’, SEQ ID NO: 3 Probe – 5’-FAM-ACACTTCTGCACTGTTAGCGGGTACA-BHQ1-3’ followed by taking into account the results using the software of the ddPCR device: calculating the absolute number of detected copies of the gene in the analysed reaction mixture with the detection of a fluorescent signal in each microdroplet.
EFFECT: invention enables detecting the SARS-CoV-2 virus regardless of the genovariation, as well as quantitative determination and dynamics of variation thereof during the different stages of the infectious process and enables making a conclusion on the clinical effectiveness on the basis of these data.
1 cl, 4 tbl, 3 ex
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