Rapid and specific identification of 5 human pathogenic Vibrio species by multiplex polymerase chain reaction targeted to dnaJ gene.

A multiplex polymerase chain reaction (PCR) method, specifically designed for application in routine diagnostic laboratories, was developed for identifying 5 human pathogen Vibrio species: Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio mimicus, and Vibrio alginolyticus. This assay directed toward the dnaJ gene was tested on a total of 355 strains representing 13 Vibrio species and 17 non-Vibrio species. Specific PCR fragments were produced in isolates belonging to the 5 target species and were absent from all strains other than these 5 species and non-Vibrio strains, indicating a high specificity of this multiplex PCR. The multiplex PCR for the detection of Vibrio pathogens in clinical specimens was experimentally applied to spiked stool samples. Only 1 specific amplicon was observed, corresponding to the pathogen spiked into the stool sample. The detection limitation was 10(5) to 10(6) cells per milliliter stool. Our data showed that this method represented a robust tool for the specific and rapid detection of the 5 major pathogenic Vibrio species.

Molecular typing of Vibrio parahaemolyticus isolates, obtained from patients involved in food poisoning outbreaks in Taiwan, by random amplified polymorphic DNA analysis.

Vibrio parahaemolyticus is one of the most important food-borne pathogens in Taiwan, Japan, and other countries with long coastlines. This paper reports on the development of a new random amplified polymorphic DNA (RAPD) method for the molecular typing of this pathogen. The 10-mer primer 284 (5'-CAG GCG CAC A-3') was selected to generate polymorphic amplification profiles of the genomic DNA at an annealing temperature of 38 degrees C. A total of 308 clinical isolates of V. parahaemolyticus collected during food poisoning outbreaks in Taiwan, mostly occurring between 1993 and 1995, plus 11 environmental and clinical reference strains were analyzed by this RAPD method. A total of 41 polymorphic RAPD patterns were recognized, and these patterns were arbitrarily grouped into 16 types (A to P). Types A, B, C, D, and E were the major types, and subtypes C3, C5, E1, B1, D2, and A2 were the major patterns. The major types were phylogenetically more closely related to each other than to any of the minor types.

[Classification of acute bacterial enteropathies]. / Inquadramento e classificazione delle enteropatie batteriche acute.

The first part considers pathogenic microorganisms (Vibrio cholerae and parahaemolytic vibrio, Clostridium welchii, enteropathogenic E. coli, Shigella, Salmonella, other enterobacteria and pseudomonas. Yersinia, simply enterotoxic Staphylococcus and that producing acute enteritis) and the process of infection (formation of a surface link without endocellular penetration with elaboration of hexotoxins, formation of a surface link with subsequent intracellular penetration, submucosa penetration). The second part discusses Salmonellae on the basis of personal experience. Particular attention is paid to current aspects of Salmonella microbiological pathomorphosis, the various isolated serotypes in relation to carriers or patients, biochemical atypias of Salmonellae strains, present-day aspects of resistance to chemoantibiotic treatment and the transfer of Salmonella Wien resistances.