Bovine leukaemia proviral DNA detection in cattle using the polymerase chain reaction.
Bovine leukaemia virus (BLV) is the causative agent in enzootic bovine leukosis a disease occurring worldwide. This virus is normally detected by the agar gel immunodiffusion or ELISA assays which rely on the appearance of antibodies to a major surface protein of the virus, gp51, present in the serum of infected cattle. We have used the polymerase chain reaction, which depends on the amplification of specific DNA sequences as a sensitive assay for the detection of BLV. It was possible to detect proviral DNA in 100 pg of tumour DNA from an infected host using agarose gel electrophoresis followed by ethidium bromide staining. The sensitivity of the assay was increased by two log orders when hybridization analysis, using a BLV proviral DNA probe, was used in combination with amplification of the DNA. Proviral DNA was detected in both lymphocytic and tumour DNA and at all stages of infection in cattle.
Naif
H M
HM
Queensland Institute of Medical Research, Herston, Brisbane, Australia.
Brandon
R B
RB
Daniel
R C
RC
Lavin
M F
MF
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D004279:DNA, Viral / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005779:Immunodiffusion; D007938:Leukemia / Q000175:diagnosis / Q000662:veterinary*; D001909:Leukemia Virus, Bovine / Q000235:genetics / Q000302:isolation & purification*; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification*
D004279:DNA, Viral; D015345:Oligonucleotide Probes; D009841:Oligonucleotides