METHOD OF DIAGNOSING WHOOPING COUGH AND DETERMINING AVIRULENT MUTANTS OF CAUSATIVE AGENT AND DIAGNOSTIC KIT Russian patent published in 2014 - IPC C12Q1/68 C07H21/02 C12P19/34 

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology and particularly to a method and diagnostic kit for diagnosing whooping cough and determining avirulent mutants of the whooping cough causative agent. The method involves carrying out a real-time polymerase chain reaction and detecting amplification products using hybridisation with fluorescent specific DNA probes. Polymerase chain reaction (PCR) is carried out using a PCR reaction mixture which comprises two pairs of primers that are suitable for amplification of fragments of sequences 1S481 and IS1002, a pair of primers for amplifying a sequence formed from insertion of IS481 and IS1002 into the cctagg site of the operon bvgAS B.pertussis, two fluorescent probes that are specific for the sequences IS481, IS1002 and the sequences formed from insertion of IS481 and IS1002 into the cctagg site of the operon bvgAS B.pertussis. The number of bacterial genones in the sample is determined from presence of sequences IS481 and IS1002 in the chromosome of B.pertussis bacteria, and the number of avirulent mutants of the causative agent is determined from presence of the insertion of IS481 and IS1002 into the cctagg site of the operon bvgAS B.pertussis. The diagnostic set comprises PCR reaction mixture, which comprises two pairs of primers that are suitable for amplification of fragments of sequences 1S481 and IS1002, a pair of primers for amplifying a sequence formed from insertion of IS481 and IS1002 into the cctagg site of the operon bvgAS B.pertussis, two fluorescent probes that are specific for the sequences IS481, IS1002 and the sequences formed from insertion of IS481 and IS1002 into the cctagg site of the operon bvg.

EFFECT: invention enables to diagnose whooping cough and determine avirulent mutants of the whooping cough causative agent.

5 cl, 1 dwg, 2 tbl, 1 ex