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United States Department of Agriculture

Agricultural Research Service

Research Project: IMPACT OF HOG AND TURKEY FARM PRODUCTION PRACTICES ON MOLECULAR EPIDEMIOLOGY OF CAMPYLOBACTER, SALMONELLA, AND EMERGING FOODBORNE PATHOGENS Title: Comparative Assessment of Standard Culture and Real-Time Pcr to Detect Campylobacter Jejuni in Retail Chicken Samples

Authors
item Debretsion, A - TUSKEGEE UNIVERSITY
item Habtemariam, T - TUSKEGEE UNIVERSITY
item Wilson, S - TUSKEGEE UNIVERSITY
item Wesley, Irene
item Yehualashet, T - TUSKEGEE UNIVERSITY

Submitted to: Journal of Food Safety
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: July 18, 2008
Publication Date: November 20, 2009
Citation: Debretsion, A., Habtemariam, T., Wilson, S., Wesley, I.V., Yehualashet, T. 2009. Comparative Assessment of Standard Culture and Real-Time PCR to Detect Campylobacter jejuni in Retail Chicken Samples. Journal of Food Safety. 29(4):588-600.

Interpretive Summary: Campylobacter is the major cause of bacterial gastroenteritis with consumption of contaminated poultry recognized as a major risk factor. We screened retail poultry carcasses for Campylobacter using a newly developed real-time PCR (rtPCR) assay and compared it to routine bacteriology. The sensitivity of the rt PCR when compared to conventional culture as the reference standard (100% sensitivity) was 81%. However, the rtPCR assay with a detection limit of 1 CFU/ml can be completed in less than three hours from sampling to final detection of C. jejuni whereas conventional culture methods require three to five days. Application of a rapid and sensitive rtPCR for detection and enumeration of C. jejuni is important for the maintenance of a safe poultry supply.

Technical Abstract: Contamination of poultry by Campylobacter is a significant source of human diarrheal illness. Conventional bacteriological methods to detect and speciate Campylobacter jejuni (C. jejuni) from chicken samples are labor-intensive and time-consuming. The purpose of this study was to compare standard culture-based methods and real-time PCR (rtPCR) for detection of C. jejuni from retail chicken samples. Culture methods (modified Campy-Cefex Agar, Charcoal cefoperazone deoxycholate agar and Brucella broth) were compared with real-time PCR (rtPCR without enrichment) for detection of C. jejuni in naturally contaminated chicken samples. Retail purchased chicken samples (n=43) were collected from four supermarkets. C. jejuni was detected by Direct Plating to Selective Agar (5/43, 11.6% DPSA), rtPCR (15/43, 34.9%), and Bolton’s enrichment (8/43, 41.9%, BE). Fifteen chicken samples were concordant by rtPCR and BE whereas three samples were positive only by BE. The sensitivity of the real-time PCR and DPSA, when compared to BE as the reference standard (100% sensitivity) were 81% and 29%, respectively. However, the rtPCR assay with a detection limit of 1 CFU/ml can be completed in less than three hours from sampling to final detection of C. jejuni. In contrast, conventional culture methods (BE and DSPA) require three to five days to speciate C. jejuni. Application of a rapid and sensitive rtPCR for detection and enumeration of C. jejuni is important for the maintenance of a safe poultry supply.

Last Modified: 8/29/2014
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