Early detection of bovine leukosis virus DNA in infected sheep using the polymerase chain reaction.
The early diagnosis of bovine leukosis virus (BLV) infection, the aetiological agent in enzootic bovine leukosis, is important for the implementation of control measures. BLV infection is currently assessed by the detection of circulating antibodies against the viral envelope protein, gp51. However, this approach has shortcomings in the time taken to detect anti-BLV antibodies (three to four weeks after infection), and in the failure to detect antibodies in some animals. Clearly a technique such as the polymerase chain reaction (PCR), which directly detects the presence of viral DNA, has advantages over methods designed to measure host antibodies. The use of PCR for the detection of proviral DNA in an affected DNA sample with as little as 10(-5) micrograms of host DNA using agarose gel electrophoresis followed by ethidium bromide staining is described here. It was possible to improve the sensitivity of this assay by using hybridisation analysis with a BLV gene probe. PCR used in combination with hybridisation analysis will provide a sensitive diagnostic assay to detect BLV when antibody tests give weakly positive or equivocal results.
Brandon
R B
RB
Molecular Oncology, Queensland Institute of Medical Research, Brisbane, Australia.
Naif
H
H
Daniel
R C
RC
Lavin
M F
MF
D000818:Animals; D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005260:Female; D007938:Leukemia / Q000175:diagnosis / Q000662:veterinary*; D001909:Leukemia Virus, Bovine / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis*
D004279:DNA, Viral; D009841:Oligonucleotides