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Construction of PCR Test-System for Differentiation between Genetically Altered Toxigenic Vibrio cholerae Strains, Biovar El Tor, with Varied Epidemic Potential

https://doi.org/10.21055/0370-1069-2014-2-85-88

Abstract

Designed is a multi-locus PCR test-system that allows for differentiation between genetically altered Vibrio cholerae strains, biovar El Tor, with high and low epidemic potential respectively, based on identification of genetic marker structure in the agent of the seventh cholera pandemic - pandemicity island VSP-II. In the course of investigations selected have been three target genes allocated in the central region and terminal end of the mobile genetic element. This test-system offers the possibility to identify the strains containing intact VSP-II, the ones containing VSP-II with a short-length deletion, and the strains with VSP-II with extended deletion. The first two are classified as the variants with low epidemic potential, while the last ones - as the variants with high epidemic potential. Specificity and efficacy of the test-system is shown by the experiments with 28 toxigenic genetically altered V. cholerae strains, biovar El Tor, and 6 strains of closely related species and enterobacteria. The results obtained coincide with the data on mono-locus PCR assay and in a number of instances are verified by sequencing.

About the Authors

D. A. Agafonov
Russian Research Anti-Plague Institute “Microbe”
Russian Federation


S. P. Zadnova
Russian Research Anti-Plague Institute “Microbe”
Russian Federation


Yu. V. Lozovsky
Russian Research Anti-Plague Institute “Microbe”
Russian Federation


N. I. Smirnova
Russian Research Anti-Plague Institute “Microbe”
Russian Federation


References

1. Zadnova S.P., Livanova L.F., Krepostnova I.M., Zakharova T.L., Osin A.V., Smirnova N.I. [Complex gene- and immune-diagnostic test-system for identification of cholera vibrios belonging to O1 and O139 serogroup; and assessment of their virulence]. RF Patent 2404257.

2. Smirnova N.I., Kirillina O.A., Cheldyshova N.B., Kutyrev V.V. [Differentiation of Vibrio cholerae eltor according to their epidemic significance using novel diagnostic cholera bacteriophages El Tor – ctx– and ctx+ and polymerase chain reaction]. Zh. Epidemiol. Mikrobiol. Immunobiol. 2001; 6:11–6.

3. Smirnova N.I., Goryaev A.A., Shubina A.V., Zadnova S.P., Kutyrev V.V. [Method of detection of toxigenic Vibrio cholerae O1 strains, identification of their biovar affinity, and differentiation of the strains belonging to biovar El Tor onto typical and genetically altered ones using multiplex polymerase chain reaction and a test-system necessary to perform the assay]. RF Patent 2458141.

4. Cherkasov A.V., Krasnov Ya.M., Agafonov D.A., Shashkova A.V., Smirnova N.I. [Variability of structure of VSP-II pandemicity island in Vibrio cholerae strains isolated in the territory of Russia]. Infektsiya i Immunitet. 2013; 3(2):185.

5. Dziejman M., Balon E., Boyd D., Fraser S.M., Heidelberg J.F., Mekalanos J.J. Comparative genomic analysis of Vibrio cholerae: Genes that correlate with cholera endemic and pandemic disease. Proc. Natl. Acad. Sci. USA. 2002; 99:1556–61.

6. Nair G.B., Qadri F., Holmgren J., Svennerholm A.M., Safa A., Bhuiyan N.A., Ahmad Q.S., Faruque S.M., Faruque A.S., Takeda Y., Sack D.A. Cholera due to altered El Tor strains of Vibrio cholerae O1 in Bangladesh. J. Clin. Microbiol. 2006; 44:4211–3.

7. O’Shea Y.A., Finnan S., Reen F.J., Morrissey J.P., O’Gara F., Boyd F. The Vibrio seventh pandemic island-II is a 26,9 kb genomic island present in Vibrio cholerae El Tor and O139 serogroup isolates that shows homology to a 43,4 kb genomic island in V. vulnificus. Microbiology. 2004; 150:4053–63.

8. Taviani E., Grim C.J., Choi J., Chun J., Haley B., Hasan N.A., Huq A., Colwell R.R. Discovery of novel Vibrio cholerae VSP-II genomic islands using comparative genomic analysis. FEMS Microbiol. Lett. 2010; 308:130–7.


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For citations:


Agafonov D.A., Zadnova S.P., Lozovsky Yu.V., Smirnova N.I. Construction of PCR Test-System for Differentiation between Genetically Altered Toxigenic Vibrio cholerae Strains, Biovar El Tor, with Varied Epidemic Potential. Problems of Particularly Dangerous Infections. 2014;(2):85-88. (In Russ.) https://doi.org/10.21055/0370-1069-2014-2-85-88

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ISSN 0370-1069 (Print)
ISSN 2658-719X (Online)