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Epidemiol Infect. 1996 Aug;117(1):59-67.

Detection of pathogenic Yersinia enterocolitica using the multiplex polymerase chain reaction.

Author information

1
Clinical Bacteriology Section, Central Public Health Laboratory, Toronto, Ontario.

Abstract

A multiplex polymerase chain reaction (PCR) was developed to detect the presence of the ail, yst, and virF genes of Yersinia enterocolitica simultaneously, quickly and accurately. The amplified fragment sizes were 356 base-pairs (bp) for the ail gene, 134 bp for the yst gene, and 231 bp for the virF gene. The specificity of the amplified products was confirmed by hybridization with digoxigenin-labelled oligonucleotide probes. Amplification was successful whether the template was derived from a single colony of bacteria, aliquots of boiled bacterial suspensions, from DNA extracted from pure or mixed cultures or from stool specimens. Amplification of the virF gene was also achieved from strains of Y. pseudotuberculosis carrying the 70 kb plasmid but not with preparations from other related Yersinia species or from other members of the family Enterobacteriaceae. The detection limit we established was 5-10 colony forming units per millilitre (cfu/ml) and 1.0 pg of DNA.

PMID:
8760951
PMCID:
PMC2271660
DOI:
10.1017/s0950268800001138
[Indexed for MEDLINE]
Free PMC Article

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