Submitted to: Food Safety Consortium Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 10/15/1999
Publication Date: N/A
Technical Abstract: In this report, we describe the development and evaluation of a 5' nuclease PCR assay for the detection of pathogenic Yersinia enterocolitica. The assay targets the chromosomally encoded invasion gene ail. Three different primer/probe sets (TM1, TM2, and TM3) amplifying different, yet overlapping, regions of ail were examined for their specificity and sensitivity. The TM1 set displayed the highest specificity, accurately detecting each of the 26 Y. enterocolitica strains tested and none of the 21 non-enterocolitica strains. This set was sensitive to approximately 0.5 pg of purified Y. enterocolitica DNA. The TM2 set was the most sensitive, allowing detection in the range of 0.25 pg of purified DNA. However, it was not specific and failed to recognize ten of the Y. enterocolitica strains used in this study. Sensitivities comparable to TM1 were achieved with the TM3 set, and cross-reaction with non-enterocolitica strains was not observed. However, this set failed to positively identify all of the Y. enterocolitica strains tested.