ALGORITHM OF TOXIGENIC GENETICALLY ALTERED VIBRIO CHOLERAE EL TOR BIOVAR STRAIN IDENTIFICATION


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Abstract

Aim. Development of an algorithm of genetically altered Vibrio cholerae biovar El Tor strain identification that ensures determination of serogroup, serovar and biovar of the studied isolate based on pheno- and genotypic properties, detection of genetically altered cholera El Tor causative agents, their differentiation by epidemic potential as well as evaluation of variability of key pathogenicity genes. Materials and methods. Complex analysis of 28 natural V. cholerae strains was carried out by using traditional microbiological methods, PCR and fragmentary sequencing. Results. An algorithm of toxigenic genetically altered V. cholerae biovar El Tor strain identification was developed that includes 4 stages: determination of serogroup, serovar and biovar based on phenotypic properties, confirmation of serogroup and biovar based on molecular-genetic properties, determination of strains as genetically altered, differentiation of genetically altered strains by their epidemic potential and detection of ctxB and tcpA key pathogenicity gene polymorphism. The algorithm is based on the use of traditional microbiological methods, PCR and sequencing of gene fragments. Conclusion. The use of the developed algorithm will increase the effectiveness of detection of genetically altered variants of the cholera El Tor causative agent, their differentiation by epidemic potential and will ensure establishment of polymorphism of genes that code key pathogenicity factors for determination of origins of the strains and possible routes of introduction of the infection.

About the authors

N. I Smirnova

Russian Research Institute of Plague Control «Microb», Saratov, Russia

D. A Agafonov

Russian Research Institute of Plague Control «Microb», Saratov, Russia

S. P Zadnova

Russian Research Institute of Plague Control «Microb», Saratov, Russia

A. V Cherkasov

Russian Research Institute of Plague Control «Microb», Saratov, Russia

V. V Kutyrev

Russian Research Institute of Plague Control «Microb», Saratov, Russia

References

  1. Методические указания «Лабораторная диагностика холеры». МУК 4.2.2218-07. М., Федеральный центр гигиены и эпидемиологии Роспотребнадзора, 2007.
  2. Смирнова Н.И., Горяев А.А., Шубина А.В. и др. Способ идентификации токсигенных штаммов V. cholerae O1, определения их биовара и дифференциации штаммов биовара эльтор на типичные и измененные методом мультиплексной полимеразной цепной реакции, и тест-система для его осуществления. Патент РФ №2458141, опубликован 05.03.2012.
  3. Шашкова А.В., Горяев А.А., Смирнова Н.И. Строение и функциональная роль CRISPR-системы бактерий. Проблемы особо опасных инфекций. 2011, 2 (108): 49-52.
  4. Chow K.H., Ng T.K., Yuen K.Y, Yam WC. Detection of RTX toxin gene in Vibrio cholerae by PCR. J. Clin. Microbiol. 2001, 39: 2594-2597.
  5. Goel A.K., Ponmariappan S., Kamboj D.V., Singh L. Single multiplex polymerase chain reaction for environmental surveillance of toxigenic-pathogenic O1 and non-O1 Vibrio cholerae. Folia Microbiol. (Praha). 2007, 52: 81-85.
  6. Grim C.J., Hasan N.A., Taviani E. et al. Genome Sequence of Hybrid Vibrio cholera O1 MJ-1236, B-33, and CIRS101 and comparative genomics with V. cholerae. J. Bacteriol. 2010, 192 (13): 3524-3533.
  7. Kumar P., Jain M., Goel A.K. et al. A large cholera outbreak due to a new cholera toxin variant of the Vibrio cholerae O1 El Tor biotype in Orissa, Eastern India. J. Med. Microbiol. 2009, 58: 234-238.
  8. Morita M., Ohnishi M., Arakawa E. et al. Development and validation of a mismatch amplification mutation PCR assay to monitor the dissemination of an emerging variant of Vibrio cholerae O1 biotype El Tor. Microbiol. Immunol. 2008, 52: 314-317.
  9. Nair G.B., Faruque S.M., Bhuiyan N.A. et al. New Variants of Vibrio cholerae O1 biotype El Tor with attributes of the classical biotype from hospitalized patients with acute diarrhea in Bangladesh. J. Clin. Microbiol. 2002, 40 (9): 3296-3299.
  10. Safa A., Nair G.B., Kong R.YC. Evolution of new variants of Vibrio cholerae O1. Trends Microbiol. 2010, 18: 46-54.
  11. Taviani E., Grim C.J., Choi J. et al. Discovery of novel Vibrio cholerae VSP-II genomic islands using comparative genomic analysis. FEMS Microbiol Lett. 2010, 308 (2): 130-137.

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Copyright (c) 2014 Smirnova N.I., Agafonov D.A., Zadnova S.P., Cherkasov A.V., Kutyrev V.V.

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