title: Development of multiplex real-time PCR assay for the detection of Brucella spp., Leptospira spp. and Campylobacter foetus.
abstract: Abortion among dairy cattle is one of the major causes of economic losses in the livestock industry. This study describes a 1-step multiplex real-time polymerase chain reaction (PCR) to detect Brucella spp., Leptospira spp. and Campylobacter foetus, these are significant bacteria commonly implicated in bovine abortion. ß-actin was added to the same PCR reaction as an internal control to detect any extraction failure or PCR inhibition. The detection limit of multiplex real-time PCR using purified DNA from cultured organisms was set to 5 fg for Leptospira spp. and C. foetus and to 50 fg for Brucella spp. The multiplex real-time PCR did not produce any non-specific amplification when tested with different strains of the 3 pathogens. This multiplex real-time PCR provides a valuable tool for diagnosis, simultaneous and rapid detection for the 3 pathogens causing abortion in bovine.
authors.affiliation: Department of Animal Medicine (Infectious Diseases), Faculty of Veterinary Medicine, Benha University, P.O. Box 13736, Toukh, Egypt.
authors.forename: Abdelfattah M
authors.initials: AM
authors.lastname: Selim
authors.forename: Mahmoud M
authors.initials: MM
authors.lastname: Elhaig
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Gaede
mesh_terms: D000818:Animals; D002002:Brucella / Q000302:isolation & purification*; D002168:Campylobacter fetus / Q000302:isolation & purification*; D002417:Cattle / Q000382:microbiology*; D007919:Leptospira / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*