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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Food Safety and Enteric Pathogens Research » Research » Publications at this Location » Publication #178950
Title: AMPLIFIED FRAGMENT LENGTH POLYMORPHISM BASED IDENTIFICATION OF GENETIC MARKERS AND NOVEL PCR ASSAY FOR DIFFERENTIATION OF CAMPYLOBACTER FETUS SUBSPECIES

Author
item VAN BERGEN, MARCEL - ANIMAL SCIENCES GROUP
item SIMONS, GUUS - KEYGENE N.V.
item VAN DER GRAAF, LINDA - ANIMAL SCIENCES GROUP
item VAN PUTTEN, JOS P - UTRECHT UNIVERSITY
item ROMBOUT, JEROEN - KEYGENE N.V.
item WESLEY, IRENE
item WAGENAAR, JAAP - ANIMAL SCIENCES GROUP

Submitted to: Journal of Medical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/9/2005
Publication Date: 12/20/2005
Citation: Van Bergen, M.A., Simons, G., Van Der Graaf, L., Van Putten, J.M., Rombout, J., Wesley, I.V., Wagenaar, J.A. 2005. Amplified fragment length polymorphism based identification of genetic markers and novel PCR assay for differentiation of campylobacter fetus subspecies. Journal of Medical Microbiology. 54(12):1217-1224.

Interpretive Summary: Campylobacter fetus subspecies venerealis (Cfv) causes infertility in cattle and is an important veterinary pathogen. Animal carriers of this bacterial agent or their sperm cannot be exported. A closely related subspecies, C. fetus subspecies fetus (Cff) causes abortion in livestock and infrequently septicemia in humans. In contrast to Cfv, animal carriers of Cff may be exported. The purpose of this study was to optimize a PCR assay specific for Cfv. Primers were based on the results of strain comparison using a highly discriminating AFLP test. The resultant assay correctly identified reference strains as well as field isolates of Cff and Cfv and was superior to currently available tests.

Technical Abstract: Subspeciation of Campylobacter fetus into C. fetus subsp. fetus (Cff) and C. fetus subsp. venerealis (Cfv) is important for both clinical and economic reasons. In the past, several molecular typing methods have been used for subspeciation, including amplified fragment length polymorphism (AFLP). In this study, we employed AFLP to identify C. fetus subspecies specific markers that can serve as a basis for design of novel PCR primer sets for Cfv. Four groups of C. fetus strains with different phenotypic or genotypic traits were examined by AFLP using 22 different DdeI/MboI primer combinations. Specific AFLP fragments were deduced and sequenced resulting in 41 sequences. Based on the obtained sequences, five potential subspecies-specific PCR assays were developed. Extensive evaluation of the five selected PCRs with a set of 65 diverse C. fetus strains identified primer set Cf C05 as subspecies Cfv specific. This newly developed PCR is fully consistent with the AFLP subspeciation results. Our data indicate AFLP as a powerful tool to compare closely related genomes and to exploit this information to develop a specific PCR with great typing potential.