Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 4/19/2000
Publication Date: N/A
Technical Abstract: We describe the development and evaluation of a fluorogenic 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 regions of ail were examined for their specificity and sensitivity. All three primer/probe sets were able to detect between 0.25 and 0.5 pg of purified Y. enterocolitica DNA. The TM1 set displayed the highest specificity detecting each of the 26 Y. enterocolitica strains tested. The TM3 set failed to identify one of the Y. enterocolitica strains, whereas the TM2 set failed to recognize 10 of the strains. None of the primer/probe sets amplified DNA from 21 non- enterocolitica strains. In addition, the TM1 set did not amplify DNA from 12 non-Yersinia pathogens commonly found in swine, indicating the assay is specific for Y. enterocolitica. Utilizing the TM1 set, the fluorogenic PCR assay was able to detect = 4 Y. enterocolitica CFU/ml in pure culture and 10 Y. enterocolitica CFU/ml independent of the presence of 10**8 CFU/ml contaminating bacteria. This set was also capable of detecting = 1 CFU of Y. enterocolitica per gram ground pork or feces after a 24-hour enrichment in ITC broth.