METHOD FOR DIFFERENTIATING TOXIGENIC AND ANTITOXIGENIC CHOLERA VIBRIOS STRAINS SEROGROUP O1 BY INHIBITING ACTIVITY Russian patent published in 2016 - IPC C12Q1/68 C12Q1/04 C12R1/63 

FIELD: medicine.

SUBSTANCE: invention relates to medical microbiology and concerns a method for differentiating toxigenic and antitoxigenic cholera vibrios strains serogroup O1 by inhibiting activity. Method involves the following steps: a) sterile tap water and cells of two toxigenic cholera vibrios strains (ctx+tcA+) and antitoxigenic (ctx-tcA-) are mixed in one vessel of 100 ml in ratio of 1:1 to the final concentration of 10⁴ m.c. per ml, the sample is incubated at 25 °C 24, for 72 or more hours; b) 0.5 ml of liquid is introduced in microtubes of 1.5 ml after incubation and DNA is recovered; c) DNA is amplified with specific primers to INDEL-gene cheA having the following sequence: cheA up-ACTTCGCGAGATTCATCAGAA; cheA dn-CTGTTTCTAGTGATGCGAAAGC, d) amplification products are analyzed by electrophoresis in 10 % polyacrylamide gel; e) obtained result is differentiated by the presence of specific alleles for each of the strains-antitoxigenic of 193 base pairs, and toxigenic -169 base pairs, visualization of presence of which makes it possible to judge on availability of inhibiting activity or its absence in the analyzed strains.

EFFECT: invention enables fast determinating ability to survive in cholera vibrios strains water bodies and can be used for monitoring water reservoirs for presence of toxigenic cholera vibrios strains and prevention of cholera.

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