title: Development of a set of multiplex standard polymerase chain reaction assays for the identification of infectious agents from aborted bovine clinical samples.
abstract: The current study describes the development of a set of 5 multiplex polymerase chain reaction (mPCR) assays for the simultaneous detection of abortive infection agents in bovine fetal tissues, including Brucella spp., Leptospira spp., and Campylobacter fetus (mPCR1); Hammondia heydorni, Neospora caninum, and Toxoplasma gondii (mPCR2); Coxiella burnetii and Chlamydophila psittaci (mPCR3); Mycoplasma bovis, Mycoplasma bovigenitalium, and Ureaplasma diversum (mPCR4); and Bovine viral diarrhea virus (BVDV) and Bovine herpesvirus-1 (BoHV-1; mPCR5). The protocol was tested on different tissue samples collected from 50 aborted bovine fetuses, and it showed that out of the 50 fetuses, 7 (14%, mPCR2) were PCR-positive for N. caninum, 4 (8%, mPCR5) were PCR-positive for BVDV, and 2 (4%, mPCR4) were PCR-positive for U. diversum. The results obtained by using each multiplex PCR were 100% concordant with those obtained by using the respective PCR assays targeting single genes on the same specimens. Moreover, all multiplex PCR assays on clinical samples were compared with reference methods, obtaining a perfect accordance in all samples and confirming the validity of the set of multiplex PCR assays. The proposed set of multiplex PCR assays is, therefore, suitable for the simultaneous detection of the main infectious agents responsible for bovine abortion.
authors: Department of Animal Production, Epidemiology and Ecology, University of Turin, Via Leonardo da Vinci, 44, 10095 Grugliasco (TO), Italy. clara.tramuta@unito.it
authors: Clara
authors: C
authors: Tramuta
authors: Daniela
authors: D
authors: Lacerenza
authors: Simona
authors: S
authors: Zoppi
authors: Mariella
authors: M
authors: Goria
authors: Alessandro
authors: A
authors: Dondo
authors: Ezio
authors: E
authors: Ferroglio
authors: Patrizia
authors: P
authors: Nebbia
authors: Sergio
authors: S
authors: Rosati
mesh_terms: D037881:Aborted Fetus / Q000382:microbiology*; D000034:Abortion, Veterinary / Q000382:microbiology*; D000818:Animals; D001419:Bacteria / Q000145:classification / Q000302:isolation & purification*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000382:microbiology* / Q000473:pathology; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity