Development and validation of a mismatch amplification mutation PCR assay to monitor the dissemination of an emerging variant of <i>Vibrio cholerae</i> O1 biotype El Tor

Morita, Masatomo; Ohnishi, Makoto; Arakawa, Eiji; Bhuiyan, N. A.; Nusrin, Suraia; Alam, Munirul; Siddique, A. K.; Qadri, Firdausi; Izumiya, Hidemasa; Nair, G. Balakrish; Watanabe, Haruo

doi:10.1111/j.1348-0421.2008.00041.x

Cited by 100 publications

(90 citation statements)

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“…This observation is interesting because the classical biotype caused the sixth pandemic before being replaced by the El Tor biotype, which is considered extinct. It was noted that altered El Tor strains replaced the prototype of the seventh pandemic El Tor in Asia (Evins et al, 1995;Nair et al, 2002) and Africa (Lizarraga-Partida et al, 2009;Morita et al, 2008). However, the Haitian ctxB variant of V. cholerae was reported to cause epidemics in Asia and Africa (Bhattacharya et al, 2016;Goel et al, 2008;Naha et al, 2012;Rashed et al, 2012;Talkington et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

“…We performed double-mismatch amplification mutation assay (DMAMA)-PCR using biotype-specific primers Rv-cla (5¢-CCTGGTACTTCTACTT-GAAACG-3¢), ctxB-3 (5¢-GTTTTACTATCTTCAGCATATGCGA-3¢) and ctxB-4 (5¢-GTTTTACTATCTTCAGCATATGCG C-3¢) for detection of classical and Haitian ctxB (Naha et al, 2012). MAMA-PCR was also performed using primers Fw-com (5¢-ACTATCTTCAGCATATG-CACATGG-3¢) and Re-elt (5¢-CCTGGTACTTCTACTTGAAACA-3¢) to determine the presence of El Tor ctxB in these isolates (Morita et al, 2008).…”

Section: Methodsmentioning

confidence: 99%

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Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India

Das

Bhotra

Zala³

et al. 2016

Journal of Medical Microbiology

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Vibrio cholerae O1 biotype El Tor, the causative agent of the seventh pandemic, has recently been replaced by strains carrying classical and Haitian ctxB in India, Haiti and other parts of the world. We conducted phenotypic and genetic tests to characterize V. cholerae O1 isolated between 2012 and 2014 from Silvassa, India, to examine the presence of virulence and regulatory genes, seventh pandemic marker, ctxB type and biofilm formation and to study genomic diversity. Of the 59 V. cholerae O1, eight isolates belong to El Tor prototype, one to classical prototype and the remaining isolates have attributes of both classical and El Tor biotypes. PCR and ctxB gene sequencing revealed the presence of classical ctxB in four strains and Haitian ctxB in 55 isolates; indicating that isolates were either an El Tor or hybrid variant. All isolates carried virulence, regulatory, adherence, Vibrio seventh pandemic pathogenicity island I and seventh pandemic group-specific marker VC2346, in addition to tcpA ET and rstR ET , the features of seventh pandemic strains, and produced cholera toxin and biofilm. PFGE analysis showed that the majority of isolates are clonal and belong to fingerprint pattern A; however, pattern B is unrelated and patterns C and D are distinct, suggesting considerable diversity in the genomic content among them. These data thus show that isolates from Silvassa are genetically diverse and that Haitian ctxB and hybrid phenotypes are undergoing global dissemination.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India

Das

Bhotra

Zala³

et al. 2016

Journal of Medical Microbiology

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“…The mismatch amplification mutation assay (MAMA) was recently developed to detect sequence polymorphism between the CL and ET ctxB genes (ctxB CL and ctxB ET , respectively) by focusing on nucleotide position 203 of the ctxB gene (Morita et al, 2008). MAMA-PCR was used to test for presence of ctxB specific for CL and ET biotypes.…”

Section: Methodsmentioning

confidence: 99%

Drug response and genetic properties of Vibrio cholerae associated with endemic cholera in north-eastern Thailand, 2003–2011

Chomvarin

Johura

Mannan

et al. 2013

Journal of Medical Microbiology

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Cholera, caused by Vibrio cholerae, results in significant morbidity and mortality worldwide, including Thailand. Representative V. cholerae strains associated with endemic cholera (n532), including strains (n53) from surface water sources, in Khon Kaen, Thailand (2003-2011, were subjected to microbiological, molecular and phylogenetic analyses. According to phenotypic and related genetic data, all tested V. cholerae strains belonged to serogroup O1, biotype El Tor (ET), Inaba (IN) or Ogawa (OG). All of the strains were sensitive to gentamicin and ciprofloxacin, while multidrug-resistant (MDR) strains showing resistance to erythromycin, tetracycline, trimethoprim/ sulfamethoxazole and ampicillin were predominant in 2007. V. cholerae strains isolated before and after 2007 were non-MDR. All except six diarrhoeal strains possessed ctxA and ctxB genes and were toxigenic altered ET, confirmed by MAMA-PCR and DNA sequencing. Year-wise data revealed that V. cholerae INET strains isolated between 2003 and 2004, plus one strain isolated in 2007, lacked the RS1 sequence (rstC) and toxin-linked cryptic plasmid (TLC)-specific genetic marker, but possessed CTX CL prophage genes ctxB CL and rstR CL . A sharp genetic transition was noted, namely the majority of V. cholerae strains in 2007 and all in 2010 and 2011 were not repressor genotype rstR CL but instead were rstR ET , and all ctx + strains possessed RS1 and TLCspecific genetic markers. DNA sequencing data revealed that strains isolated since 2007 had a mutation in the tcpA gene at amino acid position 64 (NAS). Four clonal types, mostly of environmental origin, including subtypes, reflected genetic diversity, while distinct signatures were observed for clonally related, altered ET from Thailand, Vietnam and Bangladesh, confirmed by distinct subclustering patterns observed in the PFGE (NotI)-based dendrogram, suggesting that endemic cholera is caused by V. cholerae indigenous to Khon Kaen.3These authors contributed equally to this work.Abbreviations: AMP, ampicillin; CL, classical; CTX, cholera toxin; E, erythromycin; ET, El Tor; MAMA, mismatch amplification mutation assay; MDR, multidrug resistant; SXT, trimethoprim/sulfamethoxazole; TE, tetracycline; TLC, toxin-linked cryptic plasmid. INTRODUCTIONCholera is a severe form of acute diarrhoea, caused by the gamma-proteobacterium Vibrio cholerae. V. cholerae comprises more than 200 'O' serogroups even though the disease historically has been considered to be caused only by V. cholerae serogroups O1 and O139 . Serogroup O1 has two biotypes, Classical (CL) and El Tor (ET), of which the former is associated with the first six pandemics starting in 1817 before the latter was isolated at the beginning of the current ongoing seventh cholera pandemic in the 1960s (Kaper et al., 1995). The CL biotype differs from the ET biotype in some phenotypic traits, primarily haemolysis of sheep erythrocytes, agglutination of chicken erythrocytes, Voges-Proskauer reaction, sensitivity to polymyxin B and sensitivity to specific phages (Kaper e...

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“…The strains were examined by mismatch amplifi cation mutation assay (MAMA)-based PCR for detecting the ctxB allele; a common forward primer was used for 2 alleles, FW-Com (5′-ACTATCTTCAGCATATGCACAT-GG-3′); and 2 allele-specifi c primers, Re-cla (5′-CCTGGTACTTCTAC TTGAAACG-3′) and Re-elt (5′-CCTGGTACTTCTACTTGAAA CA-3′), were used for classical and El Tor biotypes, respectively (8). Results of the MAMA-PCR are summarized in the Table. All of the 123 V. cholerae O1 strains from 1995 through 2005 yielded only the classical type of ctxB, which indicates that since 1995 the classical type has completely replaced the El Tor type ctxB (Table).…”

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confidence: 99%

ClassicalctxBinVibrio choleraeO1, Kolkata, India

Raychoudhuri¹,

Patra²,

Ghosh³

et al. 2009

Emerg. Infect. Dis.

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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

Abstract: A mismatch amplification mutation PCR assay was developed and validated for rapid detection of the biotype specific cholera toxin B subunit of V. cholerae O1. This assay will enable easy monitoring of the spread of a new emerging variant of the El Tor biotype of V. cholerae O1.

Cited by 100 publications

References 13 publications

Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India

Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India

Drug response and genetic properties of Vibrio cholerae associated with endemic cholera in north-eastern Thailand, 2003–2011

ClassicalctxBinVibrio choleraeO1, Kolkata, India

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