Study on the Development of a Rapid Detection Method for Pathogenic Yersinia enterocolitica and Yersinia pseudotuberculosis by Nested Polymerase Chain Reaction.
- Author:
Ho Jung OH
;
Hong Ki MIN
;
Yeo Won SOHN
;
Seung Hwa HONG
- Publication Type:Original Article
- MeSH:
DNA;
Limit of Detection;
Methoxsalen;
Polymerase Chain Reaction*;
Sensitivity and Specificity;
Stem Cells;
Yersinia enterocolitica*;
Yersinia pseudotuberculosis*;
Yersinia*
- From:Journal of the Korean Society for Microbiology
1999;34(2):175-187
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
A nested polymerase chain reaction (PCR) was applied to detect and identify pathogenic Yersinia enterocolitica and Y. pseudotuberculosis. We used photochemical postamplification procedure with 8-methoxypsoralen to control carryover contamination. Using the ail and inv gene, the sensitivity and specificity of DNA amplification by nested PCR was considerably improved. The amplified fragment sizes were 298 bp for the ail gene and 295 bp for the inv gene. Amplification was successful when the template was derived from three sources: purified DNA, aliquots of boiled bacterial suspension and aliquots of lysed bacterial suspension. The detection limits were 10 fg of DNA and 2 * 10 colony forming units (CFU) for Y. enterocolitica and 10 fg DNA and 2 CFU for Y. pseudotuberculosis.
