FIELD: genetic engineering; biotechnology.
SUBSTANCE: method, a specific oligonucleotide and a set for detecting hepatitis C virus RNA of genotypes 1b and 3a are proposed. A preliminary amplification of the sample material from a patient suspected of containing hepatitis C virus genotypes 1b and 3a is carried out using one or more specific oligonucleotides selected from the following SEQ ID NO: 6–11. A pre-amplified fragment of the genome of hepatitis C virus genotypes 1b and 3a is obtained. The following is prepared: a reaction mixture for detection containing the obtained target, the ribonucleoprotein complex of the CRISPR-Cas system, formed from the RNA-guided DNA endonuclease LbCpf1 from Lachnospiraceae and at least one guide RNA with the following SEQ ID NO: 1–5, fluorescent probe and detection buffer. 30–60 cycles of detection are performed in the reaction mixture in the amplifier with the detection of RNA of hepatitis C virus genotypes 1b and 3a.
EFFECT: inventions provide detection of hepatitis C virus RNA of genotypes 1b and 3a in ultra-low concentrations, up to single copies, increase in efficiency of virus RNA detection up to 5 times.
5 cl, 11 dwg, 6 tbl, 5 ex
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METHOD OF PRODUCING A PREPARATION OF A RIBONUCLEOPROTEIN COMPLEX CRISPR-CAS AND A PREPARATION FOR DETECTING HEPATITIS C VIRUS RNA OF GENOTYPES 1B AND 3A IN ULTRA-LOW CONCENTRATIONS | 2022 |
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Authors
Dates
2023-07-21—Published
2022-11-29—Filed