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J Med Microbiol 56 (2007), 56-65; DOI: 10.1099/jmm.0.46759-0
© 2007 Society for General Microbiology
ISSN 1473-5644

Rapid detection of Vibrio species using liquid microsphere arrays and real-time PCR targeting the ftsZ locus

Dobryan M. Tracz, Paul G. Backhouse, Adam B. Olson, Joanne K. McCrea, Julie A. Walsh, Lai-King Ng and Matthew W. Gilmour

National Microbiology Laboratory, Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba R3E 3R2, Canada

Correspondence
Matthew W. Gilmour
Matthew_Gilmour{at}phac-aspc.gc.ca

Received 1 June 2006
Accepted 2 October 2006

The development of rapid and sensitive molecular techniques for the detection of Vibrio species would be useful for the surveillance of sporadic infections and management of major outbreaks. Comparative sequence analysis of the ftsZ gene in the predominant Vibrio species that cause human disease revealed distinct alleles for each examined species, including Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus. Light Upon eXtension (LUX) real-time PCR assays were developed to target these species-specific polymorphisms, and were successful in rapidly differentiating the major pathogenic Vibrio species. Luminex liquid microsphere array technology was used to develop a comprehensive assay capable of simultaneously detecting V. cholerae, V. parahaemolyticus and V. vulnificus. These assays permitted the identification of a presumptive V. parahaemolyticus isolate as Vibrio alginolyticus, which was verified using additional molecular characterization.

Abbreviations: Ct, threshold cycle; LUX, Light Upon eXtension; RT-PCR, real-time PCR.

The GenBank/EMBL/DDBJ accession numbers for the ftsZ sequences are DQ520257–DQ520276.






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