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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Food Safety and Enteric Pathogens Research » Research » Publications at this Location » Publication #105676
Title: DEVELOPMENT OF A FLUOROGENIC 5' NUCLEASE PCR ASSAY FOR THE DETECTION OF PATHOGENIC YERSINIA ENTEROCOLITICA (1999 ANNUAL MEETING OF THE FOOD SAFETY CONSORTIUM)

Author
item Jourdan, Alissa
item JOHNSON, SCOTT - BIOL. RES., MILWAUKEE, WI
item Wesley, Irene

Submitted to: Food Safety Consortium Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 9/17/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: In this report, we describe the development and evaluation of a 5' nuclease PCR assay for the detection of pathogenic Yersinia enterocolitica (YE). 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 YE strains 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 10 of the YE strains used in this study. Sensitivities comparable to TM1 were achieved with the TM3 set; cross-reaction with non-enterocolitica strains was not observed. However, this set failed to positively identify all of the YE strains tested.