Abstract
Thirty-four Vibrio cholerae isolates collected from a cholera outbreak in Hyderabad, South India were found to belong to serogroup Ol biotype El Tor serotype Ogawa. The genotype of all the isolates was confirmed by PCR assays. All the isolates were found PCR positive for ctxAB, ompW, rflOl, rtxC, and tcpA genes. All the isolates but one harboured rstR El Tor allele. However, one isolate carried both rstR EL Tor as well as rstR Classical alleles. Cholera toxin (ctxB) genotyping of the isolates confirmed the presence of altered cholera toxin B of classical biotype in all the isolates. All the isolates except VCH35 harboured an RS1-CTX prophage array on the large chromosome. The isolate VCH35 contained a tandem repeat of classical CTX prophage on the small chromosome. The clonal relationship among the V. cholerae isolates as carried out by enterobacterial repetitive intergenic consensus sequences PCR, BOX PCR and randomly amplified polymorphic DNA, uniformly showed a genetic relationship among the outbreak isolates. The results of this study suggest that altered El Tor biotype V. cholerae with the classical cholera toxin gene are involved in cholera outbreaks in India.
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Ansaruzzaman, M., N.A. Bhuiyan, G.B. Nair, D.A. Sack, M. Lucas, J.L. Deen, J. Ampuero, and C.L. Chaignat. 2004. Cholera in Mozambique, variant of Vibrio cholerae. Emerg. Infect. Dis. 10, 2057–2059.
Bhattacharya, T., S. Chatterjee, D. Maiti, R.K. Bhadra, Y. Takeda, G.B. Nair, and R.K. Nandy. 2006. Molecular analysis of the rstR and orfU genes of the CTX prophages integrated in the small chromosomes of environmental Vibrio cholerae non-O1, non-O139 strains. Environ. Microbiol. 8, 526–634.
Choi, S.Y., J.H. Lee, Y.S. Jeon, H.R. Lee, E.J. Kim, M. Ansaruzzaman, N.A. Bhuiyan, and et al. 2010. Multilocus variable-number tandem repeat analysis of Vibrio cholerae O1 El Tor strains harbouring classical toxin B. J. Med. Microbiol. 59, 763–769.
Chow, K.H., T.K. Ng, K.Y. Yuen, and W.C. Yam. 2001. Detection of RTX toxin gene in Vibrio cholerae by PCR. J. Clin. Microbiol. 39, 2594–2597.
Finkelstein, R.A. 2006. Vibrio cholerae O1 and O139, and other pathogenic vibrios. [Online] http://gsbs.utmb.edu/microbook/ch024.htm.
Goel, A.K., M. Jain, P. Kumar, S. Bhadauria, D.V. Kmboj, and L. Singh. 2008. A new variant of Vibrio cholerae O1 El Tor causing cholera in India. J. Infect. 57, 280–281.
Goel, A.K., M. Jain, P. Kumar, and S.C. Jiang. 2010. Molecular characterization of Vibrio cholerae outbreak strains with altered El Tor biotype from southern India. World J. Microbiol. Biotechnol. 26, 281–287.
Goel, A.K. and S.C. Jiang. 2010. Genetic determinants of virulence, antibiogram and altered biotype among the Vibrio cholerae O1 isolates from different cholera outbreaks in India. Infect. Genet. Evol. 10, 815–819.
Goel, A.K., S. Ponmariappan, D.V. Kamboj, and L. Singh. 2007. Single multiplex polymerase chain reaction for environmental surveillance of toxigenic-pathogenic O1 and non-O1 Vibrio cholerae. Folia Microbiol (Praha). 52, 81–85.
Kaper, J.B., J.G. Morris, Jr., and M.M. Levine. 1995. Cholera. Clin. Microbiol. Rev. 8, 48–86.
Kimsey, H.H., G.B. Nair, A. Ghosh, and M.K. Waldor. 1998. Diverse CTXphis and evolution of new pathogenic Vibrio cholerae. Lancet 352, 457–458.
Kumar, P., M. Jain, A.K. Goel, S. Bhadauria, S.K. Sharma, D.V. Kamboj, L. Singh, T. Ramamurthy, and G.B. Nair. 2009. 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. 58, 234–238.
Lee, J.H., S.Y. Choi, Y.S. Jeon, H.R. Lee, E.J. Kim, B.M. Nguyen, N.T. Hien, and et al. 2009. Classification of hybrid and altered Vibrio cholerae strains by CTX prophage and RS1 element structure. J. Microbiol. 47, 783–788.
Nair, G.B., S. Misra, R.K. Bhadra, and S.C. Pal. 1987. Evaluation of the multitest medium for rapid presumptive identification of Vibrio cholerae from environmental sources. Appl. Environ. Microbiol. 53, 1203–1205.
Nair, G.B., F. Qadri, J. Holmgren, A.M. Svennerholm, A. Safa, N.A. Bhuiyan, Q.S. Ahmad, and et al. 2006. Cholera due to altered El Tor strains of Vibrio cholerae O1 in Bangladesh. J. Clin. Microbiol. 44, 4211–4213.
Nandi, B., R.K. Nandy, S. Mukhopadhyay, G.B. Nair, T. Shimada, and A.C. Ghose. 2000. Rapid method for species-specific identification of Vibrio cholerae using primers targeted to the gene of outer membrane protein OmpW. J. Clin. Microbiol. 38, 4145–4151.
Nguyen, B.M., J.H. Lee, N.T. Cuong, S.Y. Choi, N.T. Hien, D.D. Anh, H.R. Lee, and et al. 2009. Cholera outbreaks caused by an altered Vibrio cholerae O1 El Tor biotype strain producing classical cholera toxin B in Vietnam in 2007 to 2008. J. Clin. Microbiol. 47, 1568–1571.
Nusrin, S., G.Y. Khan, N.A. Bhuiyan, M. Ansaruzzaman, M.A. Hossain, A. Safa, R. Khan, and et al. 2004. Diverse CTX phages among toxigenic Vibrio cholerae O1 and O139 strains isolated between 1994 and 2002 in an area where cholera is endemic in Bangladesh. J. Clin. Microbiol. 42, 5854–5856.
Olsvik, O., J. Wahlberg, B. Petterson, M. Uhlen, T. Popovic, I.K. Wachsmuth, and P.I. Fields. 1993. Use of automated sequencing of polymerase chain reaction-generated amplicons to identify three types of cholera toxin subunit B in Vibrio cholerae O1 strains. J. Clin. Microbiol. 31, 22–25.
Pourshafie, M.R., B. Bakhshi, R. Ranjbar, M. Sedaghat, N. Sadeghifard, J. Zaemi Yazdi, M. Parzadeh, and J. Raesi. 2007. Dissemination of a single Vibrio cholerae clone in cholera outbreaks during 2005 in Iran. J. Med. Microbiol. 56, 1615–1619.
Rivera, I.G., M.A. Chowdhury, A. Huq, D. Jacobs, M.T. Martins, and R.R. Colwell. 1995. Enterobacterial repetitive intergenic consensus sequences and the PCR to generate fingerprints of genomic DNAs from Vibrio cholerae O1, O139, and non-O1 strains. Appl. Environ. Microbiol. 61, 2898–2904.
Sack, D.A., R.B. Sack, G.B. Nair, and A.K. Siddique. 2004. Cholera. Lancet 363, 223–233.
Safa, A., N.A. Bhuyian, S. Nusrin, M. Ansaruzzaman, M. Alam, T. Hamabata, Y. Takeda, D.A. Sack, and G.B. Nair. 2006. Genetic characteristics of Matlab variants of Vibrio cholerae O1 that are hybrids between classical and El Tor biotypes. J. Med. Microbiol. 55, 1563–1569.
Safa, A., G.B. Nair, and R.Y. Kong. 2010. Evolution of new variants of Vibrio cholerae O1. Trends Microbiol. 18, 46–54.
Smirnova, N.I., N.B. Cheldyshova, S.P. Zadnova, and V.V. Kutyrev. 2004. Molecular-genetic peculiarities of classical biotype Vibrio cholerae, the etiological agent of the last outbreak Asiatic cholera in Russia. Microb. Pathog. 36, 131–139.
Tamrakar, A.K., A.K. Goel, D.V. Kamboj, and L. Singh. 2006. Surveillance methodology for Vibrio cholerae in environmental samples. Int. J. Environ. Health Res. 16, 305–312.
Thiagarajah, J.R. and A.S. Verkman. 2005. New drug targets for cholera therapy. Trends Pharmacol. Sci. 26, 172–175.
Versalovic, J., M. Schneider, F.J.d. Brulin, and J.R. Lupski. 1994. Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Methods Mol. Cell. Biol. 5, 25–40.
Waldor, M.K. and J.J. Mekalanos. 1996. Lysogenic conversion by a filamentous phage encoding cholera toxin. Science 272, 1910–1914.
WHO. 2008. Cholera, 2007. Wkly Epidemiol Rec. 83, 269–283.
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Goel, A.K., Jain, M., Kumar, P. et al. Molecular characterization reveals involvement of altered el tor biotype Vibrio cholerae O1 strains in cholera outbreak at Hyderabad, India. J Microbiol. 49, 280–284 (2011). https://doi.org/10.1007/s12275-011-0317-9
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DOI: https://doi.org/10.1007/s12275-011-0317-9