Comparison of Culture, Multiplex, and 5' Nuclease Polymerase Chain Reaction Assays for the Rapid Detection of Yersinia enterocolitica in Swine and Pork Products

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Abstract:

Bacteriological culture was compared with multiplex and fluorogenic (TaqMan) polymerase chain reaction (PCR) assays for the detection of attachment invasion locus (ail)-bearing Yersinia enterocolitica in market weight swine, chitterlings, and ground pork. The TaqMan assay detected 1 pg of purified Y. enterocolitica DNA, whereas conventional gel-based PCR detected 1 ng of the same. The presence of ail-bearing Y. enterocolitica was tested in pork and feces artificially inoculated with Y. enterocolitica strain NADC 5561. The sensitivity limits of culture, multiplex, and TaqMan PCR assays were 4 × 103, 4 × 102, and 0.4 CFU/g, respectively, for the artificially inoculated pork. The sensitivity limits were 4 × 102, 4 × 102, and 0.4 CFU/g, respectively, for feces after a 48-h enrichment in a Yersinia selective broth. By the culture method, Y. enterocolitica was not detected in any of the swine specimens (n = 2,403) examined. By contrast, it was detected in 48 (2%) of the swine samples screened using the multiplex PCR and in 656 (27.2%) of these samples using the TaqMan assay. Using the culture method, Y. enterocolitica was detected in 8% of chitterling samples (n = 350) and in none of the ground pork samples (n = 350). It was identified in 27% of the chitterling samples using multiplex PCR and in 79% of these samples using the TaqMan assay. Ten percent of the ground pork samples contained Y. enterocolitica, as determined by the multiplex PCR, and 38% based on the TaqMan assay. The results suggest that pork products harbor more ail-bearing Y. enterocolitica than selected organs of freshly slaughtered hogs and that the TaqMan assay is more sensitive than either the multiplex PCR or traditional culture methods.

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Document Type: Research Article

Affiliations: 1: Pre-Harvest Food Safety and Enteric Diseases Research Unit, National Animal Disease Center, USDA, Agriculture Research Service, Ames, Iowa 50010, and Tuskegee University, Tuskegee, Alabama 36088, USA 2: Pre-Harvest Food Safety and Enteric Diseases Research Unit, National Animal Disease Center, USDA, Agriculture Research Service, Ames, Iowa 50010 3: Tuskegee University, Tuskegee, Alabama 36088, USA

Publication date: September 1, 2001

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