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Detection of pathogenic Yersinia enterocolitica using the multiplex polymerase chain reaction

Published online by Cambridge University Press:  15 May 2009

N. Harnett ,
Y. P. Lin  and
C. Krishnan
N. Harnett
Affiliation:
Clinical Bacteriology Section, Central Public Health Laboratory, Box 9000, Terminal A, Toronto, Ontario, M5W 1R5
Y. P. Lin
Affiliation:
Clinical Bacteriology Section, Central Public Health Laboratory, Box 9000, Terminal A, Toronto, Ontario, M5W 1R5
C. Krishnan
Affiliation:
Clinical Bacteriology Section, Central Public Health Laboratory, Box 9000, Terminal A, Toronto, Ontario, M5W 1R5
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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.

Type
Research Article
Information
Epidemiology & Infection , Volume 117 , Issue 1 , August 1996 , pp. 59 - 67
Copyright
Copyright © Cambridge University Press 1996

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