title: A field evaluation of the polymerase chain reaction procedure for the detection of bovine leukaemia virus proviral DNA in cattle.
abstract: A polymerase chain reaction (PCR) procedure that detects proviral bovine leukaemia virus (BLV) in peripheral blood mononuclear cell DNA was evaluated. Blood samples from all animals (164) in a commercial dairy herd with a 30% prevalence of BLV infection, and from 194 animals from BLV free herds were tested. The absence of any positive PCR results in animals from BLV free herds confirmed the specificity of the assay. Initial testing of the infected herd using a single amplification PCR (SA-PCR), detected BLV infection in 62 of 72 adult animals that were seropositive by the agar gel immunodiffusion (AGID) test and in one persistently seronegative cow. Infection in this cow was confirmed by sheep bioassay. Subsequent testing of the SA-PCR negative, seropositive animals using a double amplification PCR (DA-PCR) detected proviral BLV in eight of nine animals that were available for retesting. The PCR assay was also able to distinguish BLV infected calves from uninfected calves that were serologically positive because of the presence of colostral antibody. Lymphocytes from all seropositive animals were cultured for determination of BLV antigen expression. Cultures from 37 of 62 SA-PCR positive animals produced detectable quantities of viral antigens. However, antigen expression was not detected in cultures from seropositive animals that were negative in the SA-PCR. In addition, in experimental transmission tests, inoculation of more than 10(6) lymphocytes from these cows was required for sheep to become seropositive to BLV.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Queensland Department of Primary Industries, Animal Research Institute, Australia.
authors.forename: F W
authors.initials: FW
authors.lastname: Eaves
authors.forename: J B
authors.initials: JB
authors.lastname: Molloy
authors.forename: C K
authors.initials: CK
authors.lastname: Dimmock
authors.forename: L E
authors.initials: LE
authors.lastname: Eaves
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000097:blood* / Q000737:chemistry; D016583:Enzootic Bovine Leukosis / Q000382:microbiology* / Q000635:transmission; D005069:Evaluation Studies as Topic; D005779:Immunodiffusion; D001909:Leukemia Virus, Bovine / Q000235:genetics* / Q000302:isolation & purification; D007962:Leukocytes / Q000382:microbiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics* / Q000302:isolation & purification; D012756:Sheep
chemical_list: D017931:DNA Primers; D004279:DNA, Viral