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Applied and Environmental Microbiology, June 2003, p. 3492-3499, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3492-3499.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Rapid Detection of Campylobacter coli, C. jejuni, and Salmonella enterica on Poultry Carcasses by Using PCR-Enzyme-Linked Immunosorbent Assay

Yang Hong,1 Mark E. Berrang,2 Tongrui Liu,1 Charles L. Hofacre,1 Susan Sanchez,3 Lihua Wang,4 and John J. Maurer1,5*

Department of Avian Medicine,1 Athens Diagnostic Laboratory, College of Veterinary Medicine,3 Department of Statistics, School of Arts and Sciences, The University of Georgia, Athens, Georgia 30602,4 Agricultural Research Service, U.S. Department of Agriculture, Russell Research Center, Athens, Georgia 30604,2 Center for Food Safety and Quality Enhancement, College of Agriculture and Environmental Sciences, The University of Georgia, Griffin, Georgia 302235

Received 30 October 2002/ Accepted 10 March 2003

Contamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms require a series of biochemical and serological tests. In this study, Campylobacter ceuE and Salmonella invA genes were used to design probes in PCR-enzyme-linked immunosorbent assay (ELISA), as an alternative to conventional bacteriological methodology, for the rapid detection of Campylobacter jejuni, Campylobacter coli, and S. enterica from poultry samples. With PCR-ELISA (40 cycles), the detection limits for Salmonella and Campylobacter were 2 x 102 and 4 x 101 CFU/ml, respectively. ELISA increased the sensitivity of the conventional PCR method by 100- to 1,000-fold. DNA was extracted from carcass rinses and tetrathionate enrichments and used in PCR-ELISA for the detection of Campylobacter and S. enterica, respectively. With PCR-ELISA, Salmonella was detected in 20 of 120 (17%) chicken carcass rinses examined, without the inclusion of an enrichment step. Significant correlation was observed between PCR-ELISA and cultural methods (kappa = 0.83; chi-square test, P < 0.001) with only one false negative (1.67%) and four false positives (6.67%) when PCR-ELISA was used to screen 60 tetrathionate enrichment cultures for Salmonella. With PCR-ELISA, we observed a positive correlation between the ELISA absorbance (optical density at 405 nm) and the campylobacter cell number in carcass rinse, as determined by standard culture methods. Overall, PCR-ELISA is a rapid and cost-effective approach for the detection and enumeration of Salmonella and Campylobacter bacteria on poultry.

* Corresponding author. Mailing address: Department of Avian Medicine, College of Veterinary Medicine, The University of Georgia, Athens, GA 30603. Phone: (706) 542-5071. Fax: (706) 542-5630. E-mail: jmaurer{at}vet.uga.edu.

Applied and Environmental Microbiology, June 2003, p. 3492-3499, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3492-3499.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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