METHOD FOR DIFFERENTIATING BORDETELLA PERTUSSIS STRAINS

摘要

FIELD: medicinal microbiology, molecular biology. ^ SUBSTANCE: invention relates to molecular-genetic typing of pathogen strains of infectious diseases, detection of epidemic strains and creature perspective vaccine preparation on their base. Invention describes a method for differentiation of strains of B. pertussis by ptxS1 gene that involves isolation of DNA from strains of B. pertussis. Then isolated DNA is subjected for polymerase chain reaction with simultaneous isolation of primers S1-F2 5'-CCCCCTGCCATGGTGTGATC-3' and S1-R2 5'-AGAGCGTCTTGCGGTCGATC-3' and preparing amplicons from gene fragment encoding S1 subunit of whooping cough (ptxS1). These amplicons are subjected for successive restriction with site-specific endonucleases Ehe I and Bfu I followed by electrophoresis of restriction fragments in agarose gel and differentiation by comparison of gene ptxS1 DNA fragment amplicons size of analyzed strains of B. pertussis with control DNA samples of strains of B. pertussis comprising alleles of gene ptxS1A, ptxS1B, ptxS1D and ptxS1E. Proposed method provides precision and accelerated procedure in its carrying out in combination with economy profit. Using the method provides detection epidemic strains showing specific features of genetic structure of ptxS1 gene and to create effective vaccine preparations on their base possessing high protective activity with respect to circulating strains of B. pertussis causing the whooping cough disease. ^ EFFECT: improved method of differentiation of strains. ^ 1 ex