title: Application of a multiplex PCR assay for Campylobacter fetus detection and subspecies differentiation in uncultured samples of aborted bovine fetuses.
abstract: Campylobacter (C.) fetus (epsilonproteobacteria) is an important veterinary pathogen. This species is currently divided into C. fetus subspecies (subsp.) fetus (Cff) and C. fetus subsp. venerealis (Cfv). Cfv is the causative agent of bovine genital Campylobacteriosis, an infectious disease that leads to severe reproductive problems in cattle worldwide. Cff is a more general pathogen that causes reproductive problems mainly in sheep although cattle can also be affected. Here we describe a multiplex PCR method to detect C. fetus and differentiate between subspecies in a single step. The assay was standardized using cultured strains and successfully used to analyze the abomasal liquid of aborted bovine fetuses without any pre-enrichment step. Results of our assay were completely consistent with those of traditional bacteriological diagnostic methods. Furthermore, the multiplex PCR technique we developed may be easily adopted by any molecular diagnostic laboratory as a complementary tool for detecting C. fetus subspecies and obtaining epidemiological information about abortion events in cattle.
authors.affiliation: Evolutionary Genetics Section, Department of Animal Biology, Institute of Biology, University of the Republic, 11400 Montevideo, Uruguay.
authors.forename: Gregorio
authors.initials: G
authors.lastname: Iraola
authors.forename: Martín
authors.initials: M
authors.lastname: Hernández
authors.forename: Lucía
authors.initials: L
authors.lastname: Calleros
authors.forename: Fernando
authors.initials: F
authors.lastname: Paolicchi
authors.forename: Silvia
authors.initials: S
authors.lastname: Silveyra
authors.forename: Alejandra
authors.initials: A
authors.lastname: Velilla
authors.forename: Luis
authors.initials: L
authors.lastname: Carretto
authors.forename: Eliana
authors.initials: E
authors.lastname: Rodríguez
authors.forename: Ruben
authors.initials: R
authors.lastname: Pérez
mesh_terms: D000018:Abomasum / Q000382:microbiology; D037881:Aborted Fetus / Q000382:microbiology*; D000818:Animals; D001483:Base Sequence; D002169:Campylobacter Infections / Q000175:diagnosis / Q000662:veterinary*; D002168:Campylobacter fetus / Q000235:genetics* / Q000302:isolation & purification; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000382:microbiology*; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005819:Genetic Markers / Q000235:genetics; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011247:Pregnancy; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000662:veterinary; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D005819:Genetic Markers