|Hong, Yang - DEPT OF AVIAN MEDICINE|
|Liu, Tongrui - DEPT OF AVIAN MEDICINE|
|Hofacre, Charles - DEPT OF AVIAN MEDICINE|
|Sanchez, Susan - ATHENS DIAGNOSTICS LAB|
|Wang, Lihua - COLLEGE OF VET MED|
|Maurer, John - UGA & DEPT AVIAN MED|
Submitted to: Applied and Environmental Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: March 10, 2003
Publication Date: June 1, 2003
Citation: Hong, Y., Berrang, M.E., Liu, T., Hofacre, C.L., Sanchez, S., Wang, L., Maurer, J.J. 2003. Rapid detection of campylobacter coli, c. jejuni and salmonella enterica on poultry carcasses using pcr-elisa. Applied and Environmental Microbiology. 69(6):3492-3499. Interpretive Summary: Campylobacter and Salmonella are human pathogens that have been associated with poultry and poultry meat products. The isolation and identification of these organisms requires a series of biochemical and serological tests which can be time consuming. By combining polymerase chain reaction (PCR) and enzyme linked immuno assay (ELISA) techniques a PCR-ELISA method for the rapid detection of Campylobacter and Salmonella was developed. The method was tested using naturally contaminated poultry carcass rinse samples. The experimental methods were compared to traditional cultural bacteriological methods. The PCR-ELISA method was able to detect as few as 200 cells of Salmonella per ml and as few as 40 cells of Campylobacter per ml. Results using the PCR-ELISA method were significantly correlated with those collected with cultural methods. PCR-ELISA is a rapid and sensitive method for the detection of Campylobacter and Salmonella that may be useful in many situations where time is of the essence.
Technical Abstract: Contamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms requires a series of biochemical and serological tests. In this study, Campylobacter ceuE and Salmonella invA genes were used to design probes in PCR-ELISA, as an alternative to conventional bacteriological methodology, for the rapid detection of Campylobacter jejuni, Campylobacter coli, and Salmonella enterica from poultry samples. With PCR-ELISA (40 cycles), the detection limit for Salmonella and Campylobacter was 2 x 102 CFU/ml 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 Salmonella enterica, respectively. PCR-ELISA had an excellent, relative sensitivity (0.97) for detecting Campylobacter directly from chicken carcass rinses. With PCR-ELISA, Salmonella was detected directly from 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-squared test: p < 0.001) with only one false negative (1.67%) and four false positives (6.67%) when PCR-ELISA was used to screen sixty tetrathionate enrichment cultures for Salmonella. Overall, PCR-ELISA is a rapid and cost-effective approach for the detection of Salmonella and Campylobacter on poultry.