FIELD: biotechnology.
SUBSTANCE: nvention relates to the field of biotechnology and is a method for detecting the John Cunningham virus (JCPyV), as well as a set and specific oligonucleotide for implementing the method. First, a preliminary amplification of the sample material from a patient is carried out, a target is obtained - a preliminary amplified fragment of JCPyV using specific oligonucleotides. Next, a reaction mixture for detection is prepared, containing the specified target and the ribonucleoprotein complex of the CRISPR / Cas system, formed from the RNA-directed DNA endonuclease LbCpf1 from Lachnospiraceae, the targeting RNA, a fluorescent probe and a buffer for detection. Next, 30-60 cycles of detection are carried out in the reaction mixture in an amplifier. Invention allows efficiently detecting DNA of John Cunningham virus (JCPyV) after carrying out specific amplification of fragments of the genome of this virus.
EFFECT: ensures the detection of single copies of the DNA of the John Cunningham virus (JCPyV).
5 cl, 6 dwg, 4 tbl
Authors
Dates
2021-05-17—Published
2020-11-30—Filed