[Rapid detection and identification of enteropathogenic bacteria by a single strain counting-PCR assay].
The arbitrarily primed-PCR (AP-PCR) method is a technique to generate patterns of randomly amplified DNA fragments, and is often used for typing clinical isolates. Currently, we have been developing a modified AP-PCR method, the single strain counting-PCR (SSC-PCR), in order to discriminate among bacterial strains based on amplified patterns with multiple primers. Here, we describe the availabilities of the SSC-PCR method to identify and detect enteropathogenic bacteria. The analysis of the amplified patterns of more than 100 bacterial species shows that the patterns among strains in a species are similar to one another, and that the pattern differs among species. These results indicate that the SSC-PCR method can be a novel identification system for enteropathogenic bacteria. To examine the possibility that we can detect enteropathogenic bacteria in feces using the SSC-PCR, the model specimens were made by mixing the Vibrio parahaemolyticus DNA and the DNA directly extracted from feces. Although the patterns of fecal samples are complex with many bands, we can find a specific band of V. parahaemolyticus in the patterns of model specimens. Thus, the SSC-PCR method may be available for rapid detection of enteropathogenic bacteria in fecal specimens.
D004269:DNA, Bacterial / Q000032:analysis; D003967:Diarrhea / Q000382:microbiology*; D004755:Enterobacteriaceae / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*
D004269:DNA, Bacterial