FIELD: medicine.
SUBSTANCE: method of detection and evaluation of biotype, serogroup and toxigenicity of cholera agent with using the multipoint polymerase chain reaction (PCR) technique includes: DNA separation from the analysed object, one stage PCR production with using three pairs of the synthesised primers to a site of wbeN gene of wbf cluster, coding 01-antigen synthesis, to a site of ctxA gene coding synthesis of subunit A of cholera enterotoxin and to a site specific for comma bacillus of biovar eltor hlyA gene coding hemolysin synthesis; annealing of primers at temperature 95°-60°C during 13.5 min at number of amplification cycles 35; gel-electrophoresis; analysis of PCR results from electrophoregram and estimated results of detection and evaluation of required characteristics of comma bacillus from the presence or absence of specific strips of amplified DNA. A kit used in relation to the method comprises a complete set of reagents for DNA separation from a material, a complete set for amplification containing the primers specific to wbeN gene, hlyA gene and ctxA gene of comma bacillus, positive and negative control samples, a complete set for analysis of the products.
EFFECT: method with using the kit provides rapid, simple and precise detection of cholera agent and evaluation of its three basic properties.
3 cl, 5 dwg, 3 tbl
