Real-time PCR assay for rapid detection and quantification of Campylobacter jejuni on chicken rinses from poultry processing plant.
Campylobacter jejuni (C. jejuni) is the leading cause of food-borne gastroenteritis in the United States. Detection of Campylobacter in food samples by conventional culture is cumbersome; therefore, there is a need to develop rapid and cost-effective detection and quantification methods. Eighty-four whole chicken rinses were collected at different stages of processing at three poultry processing plants. After chicken wash collection and DNA extraction, the samples were directly subjected to real-time PCR (rtPCR) without enrichment and also culture. The assay specificity was determined with a range of Campylobacter species, related, and unrelated organisms. Of the 84 samples collected 65 (77%) of the samples were positive by the rtPCR assay and 27 (32%) of the samples tested positive by direct plating to selective agar media. The results were positively concordant for 27 (32%) of the samples. The whole rtPCR assay can be completed within 90min with a detection limit of 1CFU, compared to 5-7 days for enrichment and sub culturing in selective agar. This assay is the first report of rtPCR method capable of detecting and quantifying C. jejuni from chicken rinses without an enrichment step and could be an important, rapid and quantification model for other food-borne pathogens.
Debretsion
Aradom
A
Department of Pathobiology, College of Veterinary Medicine, Nursing & Allied Health, Tuskegee University, AL 36088, USA.
Habtemariam
Tsegaye
T
Wilson
Saul
S
Nganwa
David
D
Yehualaeshet
Teshome
T
D000818:Animals; D002138:Calibration; D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification*; D002645:Chickens / Q000382:microbiology*; D003199:Computer Systems; D005511:Food Handling; D007220:Industrial Waste / Q000032:analysis; D016133:Polymerase Chain Reaction / Q000379:methods*; D012016:Reference Values; D012680:Sensitivity and Specificity
D007220:Industrial Waste