amplified fragment length polymorphism of clinical and environmental vibrio cholerae from a freshwater environment in a cholera-endemic area, india扩增片段长度多态性的临床和环境从淡水环境中霍乱弧菌cholera-endemic地区,印度.pdfVIP

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amplified fragment length polymorphism of clinical and environmental vibrio cholerae from a freshwater environment in a cholera-endemic area, india扩增片段长度多态性的临床和环境从淡水环境中霍乱弧菌cholera-endemic地区,印度.pdf

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amplified fragment length polymorphism of clinical and environmental vibrio cholerae from a freshwater environment in a cholera-endemic area, india扩增片段长度多态性的临床和环境从淡水环境中霍乱弧菌cholera-endemic地区,印度

Mishra et al. BMC Infectious Diseases 2011, 11:249 /1471-2334/11/249 RESEARCH ARTICLE Open Access Amplified fragment length polymorphism of clinical and environmental Vibrio cholerae from a freshwater environment in a cholera-endemic area, India Arti Mishra1†, Neelam Taneja1*†, Ram K Sharma2†, Rahul Kumar2†, Naresh C Sharma3† and Meera Sharma1† Abstract Background: The region around Chandigarh in India has witnessed a resurgence of cholera. However, isolation of V. cholerae O1 from the environment is infrequent. Therefore, to study whether environmental nonO1-nonO139 isolates, which are native to the aquatic ecosystem, act as precursors for pathogenic O1 strains, their virulence potential and evolutionary relatedness was checked. Methods: V. cholerae was isolated from clinical cases of cholera and from water and plankton samples collected from freshwater bodies and cholera-affected areas. PCR analysis for the ctxA, ctxB, tcpA, toxT and toxR genes and AFLP with six primer combinations was performed on 52 isolates (13 clinical, 34 environmental and 5 reference strains). Results: All clinical and 3 environmental isolates belonged to serogroup O1 and remaining 31 environmental V. cholerae were nonO1-nonO139. Serogroup O1 isolates were ctxA, tcpA (ElTor), ctxB (Classical), toxR and toxT positive. NonO1-nonO139 isolates possessed toxR, but lacked ctxA and ctxB; only one isolate was positive for toxT and tcpA. Using AFLP, 2.08% of the V. cholerae genome was interrogated. Dendrogram analysis showed one large heterogeneous clade (n = 41), with two compact and distinct subclades (1a and 1b), and six small mono-phyletic groups. Although V. cholerae O1 isolates formed a distinct compact subclade, they were not clonal. A clinical O1 strain clustered with the nonO1-nonO139 isolates; one strain exhibited 70% similarity