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ARS Home » Southeast Area » Athens, Georgia » U.S. National Poultry Research Center » Poultry Microbiological Safety and Processing Research Unit » Research » Publications at this Location » Publication #360303

Research Project: Novel Pre-harvest Interventions and Alternatives to Antibiotics to Reduce Foodborne Pathogens

Location: Poultry Microbiological Safety and Processing Research Unit

Title: Speciation of Campylobacter spp. isolates from poultry sources with end-point PCR and real-time PCR

item Yeh, Hung-Yueh
item Plumblee Lawrence, Jodie
item Cox Jr, Nelson
item Thompson, Tori
item Berrang, Mark
item Hinton Jr, Arthur
item Brooks, Susan

Submitted to: International Poultry Scientific Forum
Publication Type: Abstract Only
Publication Acceptance Date: 12/19/2018
Publication Date: 7/15/2019
Citation: Yeh, H., Plumblee Lawrence, J.R., Cox Jr, N.A., Thompson, T.M., Berrang, M.E., Hinton Jr, A., Brooks, S.Q. 2019. Speciation of Campylobacter spp. isolates from poultry sources with end-point PCR and real-time PCR [abstract]. International Poultry Scientific Forum. 98(1):73.

Interpretive Summary: none

Technical Abstract: Statement of the purpose of the experiment: Campylobacter is the leading foodborne pathogen that causes human acute gastroenteritis worldwide. Accurate, rapid identification of Campylobacter at the species level is essential for surveillance and effective intervention. Because of their fastidious growth and sophisticated biochemical requirements, speciation of Campylobacter by traditional methods is problematic. Polymerase chain reaction (PCR) is an alternative for Campylobacter speciation. The purpose of this communication was to compare end-point PCR and BAX® System Real-Time PCR Assay for Campylobacter speciation. Description of the experimental design: Campylobacter spp., originally isolated from poultry livers in Georgia, USA (n = 250), were revived from the glycerol stocks on Campy Cefex agar plates. After 48 hours incubation under microaerobic conditions, a single colony from each isolation was cultured on fresh Campy Cefex agar plates in duplicate. Next day, colonies from one plate were examined using the BAX® System Real-Time PCR Assay, and colonies from the other plate were examined using the end-point PCR assay. Three sets of primers as previously described were used in the end-point PCR assay. Campylobacter genomic DNA used in the end-point PCR assay was isolated by boiling or by using a commercial kit. Results: The results show 181 isolates and 69 isolates were identified as Campylobacter jejuni and Campylobacter coli, respectively, from both assays. We also found that boiled DNA from seven isolates were not detected in the end-point PCR assay, but were detected using DNA templates isolated with a commercial kit, indicating that unknown inhibitors may exist in the DNA preparation with the boiling method. Conclusion: Based on our results, we concluded both PCR assays were suitable for speciation of Campylobacter isolates. Further evaluation of DNA preparation for PCR assays is needed.