Semi-nested polymerase chain reaction for detection of toxigenic Vibrio cholerae from environmental water samples

Goel, Ajay; Bhadauria, Smrati; Kumar, Pramod; Kamboj, Dev Vrat; Singh, Lokendra

doi:10.1007/s12088-007-0041-7

Cited by 5 publications

(3 citation statements)

References 19 publications

(18 reference statements)

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“…Several methods have been employed for environmental detection of V. cholerae [8][9][10][11]. However, PCR methods have their own advantages as information about presence of many toxigenic and pathogenic genes can be revealed simultaneously.…”

Section: Resultsmentioning

confidence: 99%

Characterization of Vibrio cholerae from deep ground water in a cholera endemic area in Central India

et al. 2009

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A total of 8 out of 11 deep ground water samples collected from different villages in Central India were found contaminated with Vibrio cholerae non O1, non O139. In a multiplex PCR, isolates were found positive for ompW gene but negative for ctxAB and rfbO1 genes. However, isolates from two places were positive for tcp and zot genes, indicating their intestinal colonization and toxigenic potential. Antibiotic susceptibility studies revealed that all isolates were multidrug resistant. Although, none of the isolates was found PCR positive for the mobile genetic elements, class 1 integrons and SXT constins. The results of this study corroborated that deep ground water can also be an important reservoir of V. cholerae in plane endemic areas, suggesting a continuous monitoring of water samples for timely prevention of the disease.

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Section: Resultsmentioning

confidence: 99%

Characterization of Vibrio cholerae from deep ground water in a cholera endemic area in Central India

et al. 2009

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“…All the bacterial cultures were grown at 37 °C in their respective media, procured from Difco, U.S.A and Hi-Media, India. Genomic DNA from the bacterial strains was isolated either by boiling method 26 or by DNA extraction kit (Qiagen, Hilden, Germany). 14 .…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

Development of a Polymerase Chain Reaction Assay for Detection of Burkholderia mallei, a Potent Biological Warfare Agent

et al. 2016

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Burkholderia mallei is the etiological agent of glanders, primarily a disease of equines. B. mallei is closely related to B. pseudomallei, the causative agent of melioidosis. Therefore, detection of B. mallei and its differentiation from B. pseudomallei, has always been troublesome. In present investigation, a B. mallei specific DNA sequence was identified by performing BLASTn search using ~3000 ORFs of B. mallei NCTC 10229. A polymerase chain reaction (PCR) assay with internal amplification control (IAC) was developed for detection of B. mallei and its differentiation from B. pseudomallei. The PCR assay could amplify a specific 224-bp fragment from all the six B. mallei strains used in the study, whereas other closely related organisms were tested negative. The detection limit of the assay was found to be 10 pg of purified DNA of B. mallei. Incorporation of IAC in the assay makes the results reliable as false negative results which may arise due to presence of PCR inhibitors, can be avoided. For validation, the assay was tested on tap water, Bengal gram and grass artificially spiked with B. mallei. The developed assay can be used as a simple and rapid tool for detection of B. mallei.

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“…These toxigenic and pathogenic genes have been invariably found in all the outbreak V. cholerae isolates from India [8,15]. Water, especially contaminated drinking water harbours various toxigenic V. cholerae and plays an important role in transmission of cholera [16,17].…”

Section: Distribution Of Virulent Genes Among the Isolatesmentioning

confidence: 99%

Molecular Characterization of Vibrio cholerae O1 Reveals Continuous Evolution of Its New Variants in India

et al. 2013

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Vibrio cholerae, the causing agent of cholera is still a major health challenge in most of the developing countries. In this study, V. cholerae strains collected from different cholera outbreaks in India over a period of past 7 years were found to have various toxigenic, pathogenic and regulatory genes viz. ctxAB, zot, tcp, hlyA, ace, ompU, ompW, rfbO1, toxT and toxR. The biotype specific genes rstR and rtxC revealed the El Tor biotype in majority of the isolates. However, variants among the isolates were found having genotype of both the biotypes. Sequencing of ctxB gene revealed the presence of altered ctxB of classical biotype with additional variations in isolates of 2007. Mismatch amplification mutation assay PCR also confirmed the isolates belonging to classical biotype. Antibiogram of the isolates revealed resistance for nalidixic acid, co-trimoxazole, streptomycin, and polymyxin B and susceptibility for tetracycline among most of the isolates from India. However, V. cholerae isolates from a recent outbreak in Eastern India were resistant to tetracycline. The study corroborated the continuous emergence and wide-spread of multidrug resistant El Tor variant strains in the Indian subcontinent.

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Semi-nested polymerase chain reaction for detection of toxigenic Vibrio cholerae from environmental water samples

Cited by 5 publications

References 19 publications

Characterization of Vibrio cholerae from deep ground water in a cholera endemic area in Central India

Characterization of Vibrio cholerae from deep ground water in a cholera endemic area in Central India

Development of a Polymerase Chain Reaction Assay for Detection of Burkholderia mallei, a Potent Biological Warfare Agent

Molecular Characterization of Vibrio cholerae O1 Reveals Continuous Evolution of Its New Variants in India

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