Comparison of culture, multiplex, and 5' nuclease polymerase chain reaction assays for the rapid detection of Yersinia enterocolitica in swine and pork products.
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 I 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 x 10(3), 4 x 10(2), and 0.4 CFU/g, respectively, for the artificially inoculated pork. The sensitivity limits were 4 x 10(2), 4 x 10(2), 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.
Boyapalle
S
S
Pre-Harvest Food Safety and Enteric Diseases Research Unit, National Animal Disease Center, USDA, Agriculture Research Service, Ames, Iowa 50010, USA.
Wesley
I V
IV
Hurd
H S
HS
Reddy
P G
PG
D000818:Animals; D015373:Bacterial Typing Techniques; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis*; D004768:Enterotoxins; D005243:Feces / Q000382:microbiology; D005456:Fluorescent Dyes; D008461:Meat Products / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013552:Swine; D019914:Taq Polymerase; D015008:Yersinia enterocolitica / Q000235:genetics* / Q000302:isolation & purification
D004269:DNA, Bacterial; D004768:Enterotoxins; D005456:Fluorescent Dyes; D019914:Taq Polymerase