[Toxigenic Vibrio cholerae: identification of the ctxB gene]

A specific probe was designed to identify part of the genetic sequence of the ctxB gene which encodes for the B subunit of the cholera toxin by polymerase chain reaction (PCR) which amplifies a 318 bp segment of the ctxB gene. Marked with P32, we used this probe for colony hybridization which is a technique for identifying the production capacity of subunit B of strains of Vibrio cholerae O1 from different outbreaks in South America (Perú 1992 and Ecuador 1993-1995) and from, collection strains. This probe was tested for the identification of the ctxB gene in Vibrio cholerae O139.

Thirty-eight phylogenetically related strains were studied: 24 V. cholerae O1, 4 V. cholerae non O1, 5 Aeromonas, 4 Plesiomonas and 1 Escherichia coli.

The probe demonstrated to be useful for the identification of the ctxB gene (which codifies for the subunit B of the cholera toxin) in 24 strains of Vibrio cholerae O1 and in the Vibrio cholerae O139 strain. The ctxB gene was not detected in the remaining strains pertaining to the Vibrio cholerae non O1 species (non O139), Plesiomonas, Aeromonas spp. and E. coli. The specificity of this product was not demonstrated since no signal of unspecific hybridization appeared with phylogenetically related strains such as Escherichia coli K88 (LT+) and Aeromonas hydrophila ATCC (LT+), producers of the thermolabile LT toxin. It is important to indicate that the ctxB gene in V. cholerae O139 has been identified, for the first time, with our probe and thus it may be said that all the strains which have genetic codification for CT up to now may be identified.

We conclude that the system herein described provides advantages over the immunologic and biologic methods for evaluating a large number of samples in a short time and with excellent specificity and sensitivity which are important in the diagnosis and the epidemiologic surveillance of the disease.