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The Journal of Medical Microbiology, Vol 48, Issue 10 883-889, Copyright © 1999 by Society for General Microbiology

JOURNAL ARTICLE

An improved method for detecting faecal Vibrio cholerae by PCR of the toxin A gene

K. Miyagi, K. Sano, C. Morita, S. Imura, S. Morimatsu, T. Goto, Y. Nakano, K. Omura, Y. Matsumoto, K. Maeda, S. Hashimoto and T. Honda
Department of Microbiology, Osaka Medical College, Japan. ksano@art.osaka-med.ac.jp

A method for removing inhibitor(s) of the PCR assay for the direct detection of cholera toxin A gene (ctxA) in human faeces is described. Inhibitors of the PCR were removed by centrifugation and the activity of the remaining inhibitors by dilution. Based on these data, a protocol was developed for pre-treatment of stool specimens for PCR assay, and a simple and rapid protocol was constructed for the diagnostic detection of the ctxA genes in stool specimens in combination with single band detection on gel electrophoresis, dot-blot hybridisation and enrichment culture. This protocol was applied to clinical specimens and showed that the PCR method gave 100% agreement with established culture methods for the detection of cholera toxin-producing Vibrio cholerae O1. This protocol was considered to be useful because of its simplicity and the rapidity of diagnosis.


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[Abstract] [Full Text]


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