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

A method for removing inhibitor(s) of the PCR assay for the direct detectio n of cholera toxin A gene (ctxA) in human faeces is described. Inhibitors o f the PCR were removed by centrifugation and the activity of the remaining inhibitors by dilution. Based on these data, a protocol was developed for p re-treatment of stool specimens for PCR assay, and a simple and rapid proto col 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 wi th established culture methods for the detection of cholera toxin-producing Vibrio cholerae O1. This protocol was considered to be useful because of i ts simplicity and the rapidity of diagnosis.