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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 #115522
Title: SENSITIVITY LIMITS OF CULTURE, CONVENTIONAL AND FLUOROGENIC PCR ASSAYS IN THE DETECTION OF AIL-BEARING YERSINIA ENTEROCOLITICA

Author
item BOYAPALLE, S - 3625-30-15
item Wesley, Irene

Submitted to: Food Safety Consortium Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 9/19/2000
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Bacteriological culture was compared with polymerase chain reaction (PCR)-based protocols, including conventional and fluorogenic 5' nuclease PCR assays for the detection of ail-bearing Y. enterocolitica. The presence of Y. enterocolitica was tested in pure cultures and in irradiated pork and feces inoculated with Y. enterocolitica strain NADC 5561. Fluorogenic PCR assay was able to detect < 1 pg of purified Y. enterocolitica DNA; whereas conventional gel-based PCR was able to detect > 1 ng of the same. The sensitivity limits of culture, conventional and fluorogenic PCR assays to detect Y. enterocolitica were 4 x 10**3, 4 x 10**2, and 0.4 CFU/g, respectively, for the pork inoculated with known concentrations of the strain NADC 5561. The sensitivity limits were 4 x 10**2, 4x10**2, and 0.4 CFU/g, respectively, for feces. The results suggest that the fluorogenic PCR assay is more sensitive at detecting ail- bearing Y. enterocolitica than either conventional PCR or traditional culture methods.