Multiplex PCR for the differentiation of Vibrio cholerae 01 biovar El Tor strains of different epidemic importance


DOI: https://dx.doi.org/10.18565/epidem.2018.4.49-55

Agafonova E.Yu., Agafonov D.A., Smirnova N.I.

Russian Research Institute «Microbe», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Saratov, Russia
Objective. To develop a procedure for the simultaneous identification of toxic epidemically dangerous Vibrio cholerae biovar El Tor strains and for the differentiation of nontoxigenic strains into potentially epidemic dangerous and epidemic safe ones.
Materials and methods. The investigators used 167 V. cholerae O1 biovar El Tor strains, 6 nontoxigenic V. cholerae non-O1/O139 strains, 5 bacterial strains of closely related species in the genus Vibrio, and 3 Enterobacteriaceae strains.
Results. One-step multiplex PCR for the identification of marker genes was carried out in 2 reaction mixtures. The toxigenic epidemically dangerous strains were typified by the formation of 4 amplified fragments corresponding to the vc0180, vc0514, ctxA, and tcpA genes; the nontoxigenic potentially epidemically dangerous strains were characterized by 3 fragments corresponding to the vc0180, vc0514 and tcpA genes, and the nontoxigenic epidemically safe strains were typical of the formation of 1 fragment corresponding to the tcpA gene, or the absence of all detected marker genes, respectively.
Conclusion. A procedure has been developed to differentiate V, cholerae O1 biovar El Tor strains of different epidemic importance. The investigation has shown the specificity and effectiveness of the developed multiplex PCR that can be used in epidemiological investigations, monitoring studies of environmental objects and for the certification of V. cholera strains.
Keywords: epidemic importance, nontoxigenic strains, Vibrio cholerae biovar El Tor, polymerase chain reaction
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For citations: Agafonova E.Yu., Agafonov D.A., Smirnova N.I. Multiplex PCR for the differentiation of Vibrio cholerae 01 biovar El Tor strains of different epidemic importance. Èpidemiologiâ i infekcionnye bolezni. Аktual’nye voprosy 2018; (4):49–55 (In Russ.).


About the Autors

Elena Yu. Agafonova, Junior Reasecher, Department of Microbiology, Russian Research Institute «Microbe» №, Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Saratov, Russia; е-mail: elenabaranichina@ mail.ru; ORCID: https://orcid.org/0000-0001-9988-6312
Dmitriу A. Agafonov, Cand. Biol. Sci., Senior Researcher, Department of Microbiology, Russian Research Institute «Microbe», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Saratov, Russia; e-mail: rusrapi@microbe.ru; ORCID: https://orcid.org/0000-0001-9273-6063
Prof. Nina I. Smirnova, Biology Dr; Head, Department of Microbiology, Russian Research Institute «Microbe», Russian Federal Service for Supervision of Consumer Rights Protection and Human Well-Being, Saratov, Russia; e-mail: rusrapi@microbe.ru; ORCID: https://orcid.org/0000-0002-7115-6286


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