start-ver=1.4
cd-journal=joma
no-vol=14
cd-vols=
no-issue=11
article-no=
start-page=e02661-25
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2026
dt-pub=20261103
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=A novel SNP-based multiplex PCR profiling divulges contemporary genetic patterns in Haitian variant Vibrio cholerae O1 clinical isolates
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=The massive cholera outbreak in Haiti in 2010 has drawn the attention of both medical professionals and public health officials worldwide. The hypervirulent Vibrio cholerae O1 El Tor strains responsible for this outbreak were not only confined to Haiti but also spread quickly to other cholera-prone regions. Therefore, tracking and understanding these strains in endemic areas have become crucial for timely and effective treatment strategies. The presence of specific genetic mutations in key virulence-related genes—ctxB (cholera enterotoxin subunit B), tcpA (toxin-coregulated pilin), and rtxA (repeats-in-toxin)—makes these Haitian strains distinctive. These genes contain single nucleotide polymorphisms, which act as unique markers for identifying the Haitian variant. However, current detection methods using allele-specific simplex PCRs are time-consuming and can delay critical outbreak responses. To address this, we developed new Haitian allele-specific primers targeting rtxA and tcpA and combined them with an existing ctxB-specific PCR into a single multiplex PCR assay. We tested this new multiplex PCR on 250 clinical V. cholerae O1 isolates collected from diarrheal patients in Kolkata, India, between 2002 and 2019. Interestingly, we noticed that the first appearance of Haitian tcpA, rtxA, and ctxB alleles was in 2003, 2004, and 2006, respectively, years before the Haitian outbreak occurred. Our results matched perfectly with the conventional simplex PCRs and demonstrated 100% sensitivity and specificity. Given the global spread of these Haitian-like V. cholerae strains, this multiplex PCR provides a fast, reliable tool for identifying cholera infection and gaining valuable epidemiological insights.
en-copyright=
kn-copyright=

en-aut-name=SamantaProsenjit
en-aut-sei=Samanta
en-aut-mei=Prosenjit
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=

en-aut-name=GhoshPriyanka
en-aut-sei=Ghosh
en-aut-mei=Priyanka
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=

en-aut-name=ChowdhuryGoutam
en-aut-sei=Chowdhury
en-aut-mei=Goutam
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=

en-aut-name=ShawSreeja
en-aut-sei=Shaw
en-aut-mei=Sreeja
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=

en-aut-name=NahaArindam
en-aut-sei=Naha
en-aut-mei=Arindam
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=

en-aut-name=RoyDeboleena
en-aut-sei=Roy
en-aut-mei=Deboleena
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=

en-aut-name=OhnoAyumu
en-aut-sei=Ohno
en-aut-mei=Ayumu
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=

en-aut-name=MiyoshiShin-ichi
en-aut-sei=Miyoshi
en-aut-mei=Shin-ichi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=

en-aut-name=RamamurthyThandavarayan
en-aut-sei=Ramamurthy
en-aut-mei=Thandavarayan
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=

en-aut-name=MukhopadhyayAsish Kumar
en-aut-sei=Mukhopadhyay
en-aut-mei=Asish Kumar
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=10
ORCID=

affil-num=1
en-affil=Division of Bacteriology, ICMR-National Institute for Research in Bacterial Infections
kn-affil=

affil-num=2
en-affil=Division of Bacteriology, ICMR-National Institute for Research in Bacterial Infections
kn-affil=

affil-num=3
en-affil=Collaborative Research Centre of Okayama University for Infectious Diseases, ICMR-National Institute for Research in Bacterial Infections
kn-affil=

affil-num=4
en-affil=Division of Bacteriology, ICMR-National Institute for Research in Bacterial Infections
kn-affil=

affil-num=5
en-affil=Division of Bacteriology, ICMR-National Institute for Research in Bacterial Infections
kn-affil=

affil-num=6
en-affil=Division of Bacteriology, ICMR-National Institute for Research in Bacterial Infections
kn-affil=

affil-num=7
en-affil=Collaborative Research Centre of Okayama University for Infectious Diseases, ICMR-National Institute for Research in Bacterial Infections
kn-affil=

affil-num=8
en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
kn-affil=

affil-num=9
en-affil=Division of Bacteriology, ICMR-National Institute for Research in Bacterial Infections
kn-affil=

affil-num=10
en-affil=Division of Bacteriology, ICMR-National Institute for Research in Bacterial Infections
kn-affil=
en-keyword=cholera
kn-keyword=cholera
en-keyword=Vibrio cholerae
kn-keyword=Vibrio cholerae
en-keyword=multiplex PCR
kn-keyword=multiplex PCR
en-keyword=ctxB
kn-keyword=ctxB
en-keyword=tcpA
kn-keyword=tcpA
en-keyword=rtxA
kn-keyword=rtxA
END