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

被引：6

作者：

Goel A.K. ^{[1
]}

Bhadauria S. ^{[1
]}

Kumar P. ^{[1
]}

Kamboj D.V. ^{[1
]}

Singh L. ^{[1
]}

机构：

[1] Biotechnology Division, Defence Research and Development Establishment

来源：

Indian Journal of Microbiology | 2007年 / 47卷 / 3期

关键词：

Environmental detection; Semi-nested PCR; Toxigenic; Viable; Vibrio cholerae;

D O I：

10.1007/s12088-007-0041-7

中图分类号：

学科分类号：

摘要：

A rapid and sensitive direct cell semi-nested PCR assay was developed for the detection of viable toxigenic V. cholerae in environmental water samples. The semi-nested PCR assay amplified cholera toxin (ctxA2B) gene present in the toxigenic V. cholerae. The detection sensitivity of direct cell semi-nested PCR was 2 × 103 CFU of V. cholerae whereas direct cell single-step PCR could detect 2 × 104 CFU of V. cholerae. The performance of the assay was evaluated using environmental water samples after spiking with known number of Vibrio cholerae O1. The spiked water samples were filtered through a 0.22 micrometer membrane and the bacteria retained on filters were enriched in alkaline peptone water and then used directly in the PCR assay. The semi-nested PCR procedure coupled with enrichment could detect less than 1 CFU/ml in ground water and sea water whereas 2 CFU/ml and 20 CFU/ml could be detected in pond water and tap water, respectively. The proposed method is simple, faster than the conventional detection assays and can be used for screening of drinking water or environmental water samples for the presence of toxigenic V. cholerae. © 2007 Association of Microbiologists of India.

引用

页码：207 / 211

页数：4

共 18 条

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