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Year : 2016  |  Volume : 144  |  Issue : 2  |  Page : 293-296

Haitian variant tcpA in Vibrio cholerae O1 El Tor strains in National Capital Region (India)

1 Laboratory Department, Maharishi Valmiki Infectious Diseases Hospital, New Delhi, India
2 National Institute of Cholera & Enteric Diseases, Kolkata, West Bengal, India

Date of Submission17-Jan-2014
Date of Web Publication1-Dec-2016

Correspondence Address:
Naresh Chand Sharma
Laboratory Department, Maharishi Valmiki Infectious Diseases Hospital, New Delhi
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0971-5916.195054

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How to cite this article:
Kumar D, Sharma NC, Ramamurthy T, Gupta SK, Seth DK. Haitian variant tcpA in Vibrio cholerae O1 El Tor strains in National Capital Region (India). Indian J Med Res 2016;144:293-6

How to cite this URL:
Kumar D, Sharma NC, Ramamurthy T, Gupta SK, Seth DK. Haitian variant tcpA in Vibrio cholerae O1 El Tor strains in National Capital Region (India). Indian J Med Res [serial online] 2016 [cited 2020 Jun 4];144:293-6. Available from:


Vibrio cholerae O1 is the causative agent of cholera, which has two biotypes, namely, classical and El Tor, based on various phenotypic and genotypic characters [1] . Cholera toxin (ctx) and toxin co-regulated pilus (tcp) are essential virulence genes. The expression of CT and TCP is regulated by ToxR, a co-regulatory protein [2] . TCP is a type IV pilus which is essential for colonization in the small intestine [3] . Whole genome sequence of V. cholerae strains isolated from Bangladesh (CIRS101) and Haitian outbreak has shown a single nucleotide polymorphism (SNP) at a nucleotide position 266 (amino acid 89) of the tcpA gene, particularly associated with the Haitian variant [4],[5],[6] . We undertook this study to investigate the presence of mutation in tcpA allele in V. cholerae El Tor strains obtained from National Capital Region (NCR) of India.

A total of 71 V. cholerae strains could be revived from the collection maintained in the Laboratory Department of Maharishi Valmiki Infectious Diseases Hospital (MVIDH), Delhi, India. These strains were collected from NCR (Delhi, Haryana and Uttar Pradesh) during 2008-2012 and were characterized biochemically and serotyped as V. cholerae O1 Ogawa using standard procedure [7] . Environmental sampling and processing of samples were done according to Mishra et al[8] . A total of 204 samples were collected which included 171 drinking water samples from the houses of patients admitted to MVIDH, 13 samples from Najafgarh drain, seven samples from different lakes and three samples from the Yamuna river. Only two drinking water samples and one sewage samples were positive for V. cholerae O1. Detection of the mutation in tcpA gene encoding TCP was done by a polymerase chain reaction (PCR) assay [9] . This PCR assay discriminates the V. cholerae strains harbouring Haitian, classical and El Tor alleles of tcpA, and this may be used to understand the presence of the new variant in different areas of cholera endemicity.

In this study, three different primers used included one common reverse primer for both El Tor and Haitian type tcpA alleles [tcpA El-Rev (5'-CCGACTGTAATTGCGAATGC-3')]. Two forward primers [tcpA-F'1 (5'-CCAGCTACCGCAAACGCAGA-3') and tcpA-F'2 (5'-CCAGCTACCGCAAACGCAGG-3')] specific for El Tor and Haitian type tcpA alleles were used, respectively. The PCR assay conditions and PCR cycles were as described previously [9] . N16961 was used as a control strain for El Tor, and EL-1786 for Haitian, to check the mutation in tcpA gene. These control strains were obtained from National Institute of Cholera and Enteric Diseases (NICED), Kolkata, India.

PCR assay confirmed all the 71 strains carrying tcpA of Haitian type which yielded a 167bp fragment with Haitian-specific primer pair but not with El Tor-specific primer. Only V. cholerae O1 Inaba El Tor biotype (control strain N16961) was amplified with El Tor-specific primers which yielded a 167bp fragment but not with Haitian tcpA-specific primer. Previous studies reported a single nucleotide change at 266 position which resulted in asparagine to serine substitution [4],[9] . This mutation (Asn→Ser) at the 89 th amino acid of whole tcpA was the result of transition which took place in isolates of Bangladesh in 2002 [10] and isolates of Kolkata, India, in 2003 [9] . Haitian tcpA allele (tcpETCIRS ) has been found among the isolates of Afghanistan, Cameroon, India, Nepal, Nigeria, Pakistan, South Africa and Sri Lanka [5] . In this study, the combination of Haitian ctxB (ctxB7), classical ctxB (ctxB1) and tcpA of Haitian allele is reported for the first time since 2008 in north India including Delhi, Haryana and Uttar Pradesh in both 69 clinical and two environmental isolates (sewage & drinking water) [Table 1]. The presence of tcpA Haitian allele was not restricted to strains having ctxB7 allele. It was equally present among both ctxB1 and ctxB7 isolates. Such molecular intricacies are important to understand the contemporary developments taking place in ctxB and tcpA genes globally. Keeping in view of the Haiti experiences and findings in our country, there is a need for the redressal of control strategies being adopted in the surveillance of cholera disease in this endemic region.
Table 1. The list of clinical (n=69) and environmental (n=2) Vibrio cholerae strains based on polymerase chain reaction study isolated from diarrhoeal patients in north India

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   References Top

Morita M, Ohnishi M, Arakawa E, Bhuiyan NA, Nusrin S, Alam M, 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 : 314-7.  Back to cited text no. 1
Faruque SM, Albert MJ, Mekalanos JJ. Epidemiology, genetics, and ecology of toxigenic Vibrio cholerae. Microbiol Mol Biol Rev 1998; 62: 1301-14.  Back to cited text no. 2
Herrington DA, Hall RH, Losonsky G, Mekalanos JJ, Taylor RK, Levine MM. Toxin, toxin-coregulated pili, and the toxR regulon are essential for Vibrio cholerae pathogenesis in humans. J Exp Med 1988; 168 : 1487-92.  Back to cited text no. 3
Grim CJ, Hasan NA, Taviani E, Haley B, Chun J, Brettin TS, et al. Genome sequence of hybrid Vibrio cholerae O1 MJ-1236, B-33, and CIRS101 and comparative genomics with V. cholerae. J Bacteriol 2010; 192: 3524-33.  Back to cited text no. 4
Talkington D, Bopp C, Tarr C, Parsons MB, Dahourou G, Freeman M, et al. Characterization of toxigenic Vibrio cholerae from Haiti, 2010-2011. Emerg Infect Dis 2011; 17 : 2122-9.  Back to cited text no. 5
Chin CS, Sorenson J, Harris JB, Robins WP, Charles RC, Jean-Charles RR, et al. The origin of the Haitian cholera outbreak strain. N Engl J Med 2011; 364 : 33-42.  Back to cited text no. 6
World Health Organization. Manual for the Laboratory Identification and Antimicrobial Susceptibility Testing of Bacterial Pathogens of Public Health Importance in the Developing World. Centers for Disease Control and Prevention, Atlanta Georgia USA. USAID/WHO/CDC; 2003. p. 141-59.  Back to cited text no. 7
Mishra A, Taneja N, Sharma RK, Kumar R, Sharma NC, Sharma M. Amplified fragment length polymorphism of clinical and environmental Vibrio cholerae from a freshwater environment in a cholera-endemic area, India. BMC Infect Dis 2011; 11: 249.  Back to cited text no. 8
Ghosh P, Naha A, Basak S, Ghosh S, Ramamurthy T, Koley H, et al. Haitian variant tcpA in Vibrio cholerae O1 El Tor strains in Kolkata, India. J Clin Microbiol 2014; 52 : 1020-1.  Back to cited text no. 9
Hasan NA, Choi SY, Eppinger M, Clark PW, Chen A, Alam M, et al. Genomic diversity of 2010 Haitian cholera outbreak strains. Proc Natl Acad Sci USA 2012; 109 : E2010-7.  Back to cited text no. 10


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