SET OF OLIGONUCLEOTIDE PRIMERS FOR TYPING OF HISTOPLASMA CAPSULATUM STRAIN HISTOPLASMOSIS AGENT BY AMPLIFICATION OF DIFFERENTIATING DNA FRAGMENTS (DFR) Russian patent published in 2018 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention can be used for intraspecific differentiation of Histoplasma capsulatum strains by specialists of reference centres for infectious agents monitoring, as well as research organizations of epidemiological and microbiological profile. This invention is intended to provide a set of oligonucleotide primers for intraspecific typing of H. capsulatum micromycetes by amplification of differentiating regions of genomic DNA. The goal is achieved by construction of a set of specific oligonucleotides for intraspecific typing of histoplasmosis agent strains by amplifying 7 differentiating DNA fragments of H. capsulatum having direct and reverse primer activity in the amplification reaction with the following structure: 5' -CTTCTTATTCCTTGTATCAC -3' - HcDFR 3s, 5' -CATCTTATTGTCACTGTTTC -3' - HcDFR 3as for amplification of a differentiating fragment of HcDFR3 with a length of 504 bps; 5' -GAGTTTCTTACATCAATACA -3' - HcDFR 7s 5' -CTCTGCTTCTACTTATCTG -3' - HcDFR 7as for amplification of a differentiating fragment of HcDFR7 with a length of 199 bps; 5' -GAAGCAGAAGCAGTATGAGG -3' -HcDFR 8s, 5' -TACGGACAATGACAAATGAG -3' -HcDFR 8as for amplification of a differentiating fragment of HcDFR8 with a length of 172 bps; 5' -GAAGCAGAAGCAGTATGAGG -3' - HcDFR 9s, 5' -CAATGACAAATGAGCAGG -3' - HcDFR 9as for amplification of a differentiating fragment of HcDFR9 with a length of 166 bps; 5' -CAGTTGGATAGATGATGT -3' - HcDFR 10s, 5' -ATGAAGAAGGAGTTGAG -3' - HcDFR Was for amplification of a differentiating fragment of HcDFRIO with a length of 188 bps; 5' -GAAGCAGAAGCAGTATGAGG -V-HcDFR 11s, 5' -ATACGGACAATGACAAATGA -3' - HcDFR 11as for amplification of a differentiating fragment of HcDFR11 with a length of 173 bps; 5' -CGGACAATGACAAATGA -3' - HcDFR 12s, 5' -CAAACAGCAGATAAAGACAA -3' - HcDFR 12as for amplification of a differentiating fragment of HcDFR12 with a length of 134 bps. The developed set of oligonucleotide primers for Histoplasma capsulatum histoplasmosis strains typing by amplification of differentiating DNA fragments allows intraspecific genetic typing of H. capsulatum strains with high reproducibility and resolution, as well as relatively low cost. The proposed method uses only equipment for PCR with appropriate reagents.

EFFECT: possibility of widespread implementation of this molecular genetic approach in research organizations and specialized laboratories of epidemiological and microbiological profile.

2 dwg, 2 tbl, 3 ex