The polymerase chain reaction (PCR) was used to detect bovine leukemia virus in bovine blood samples. When applied to leucocytes extracted from the blood samples, the standard method of DNA extraction gave good correlation with agar gel immunodiffusion, but a method in which 5 μl of blood was the starting material was unreliable. Selection of the primers was important, and differences in results were observed when the PCR method was applied to blood samples from different geographic areas. The sensitivity varied from 50% to 90%, depending on the primer set applied to the gag gene of proviral nucleic acid. This variation was based on geographic origin of the cattle, suggesting an influence of viral strain. In some areas, more than 1 primer may needed to optimize results.
References
1.
AgrestiAPontiWRocchiM: 1993, Use of polymerase chain reaction to diagnose bovine leukemia virus infection in calves at birth. Am J Vet Res3:373–378.
2.
Ballagi-PordanyAKlintevallKMerzaM: 1992, Direct detection of bovine leukemia virus infection: practical applicability of a double polymerase chain reaction. J Vet Med Ser B39:69–77.
3.
BrandonRBNaifHDanielRCLavinMF: 1991, Early detection of bovine leukosis virus DNA in infected sheep using the polymerase chain reaction. Res Vet Sci50:89–94.
4.
BumyABruckCChantrenneH: 1980, Bovine leukaemia virus: molecular biology and epidemiology. In: Viral epidemiology and oncology, ed. KleinG, pp. 231–289. Raven Press, New York, NY.
5.
CoulstonJNaifHBrandonR: 1990, Molecular cloning and sequencing of an Australian isolate of proviral bovine leukaemia virus DNA: comparison with other isolates. J Gen Virol71:1737–1746.
6.
GravesDCDiglioCAFerrerJF: 1977, A reverse transcriptase assay for detection of the bovine leukemia virus. Am J Vet Res38:1739–1744.
7.
HavePHoff-JorgensenR: 1991, Demonstration of antibodies against bovine leukemia virus (BLV) by blocking ELISA using polyclonal anti-BLV immunoglobulin. Vet Microbiol27:221–229.
8.
HouseJAGloverFLHouseC: 1975, Current aspect of bovine leukemia. Proc Annu Conv Am Assoc Bovine Pract8:147–150.
9.
JacobsRMSongZPoonH: 1992, Proviral detection and serology in bovine leukemia virus-exposed normal cattle and cattle with lymphoma. Can J Vet Res56:339–348.
10.
KellyEJJacksonMKMarsolaisG: 1993, Early detection of bovine leukemia virus in cattle by use of the polymerase chain reaction. Am J Vet Res54:205–209.
11.
KettmannRCleuterYMammerickxM: 1980, Genomic integration of bovine leukemia virus: comparison of persistent lymphocytes with lymph node tumor forms of enzootic bovine leukosis. Proc Natl Acad Sci USA77:2577–2481.
12.
KuzmakJSkorupskaAMoussaAGrundboeckJ: 1993, Application of non-radioactive method of DNA detection in the diagnosis of bovine leukaemia virus infection. Bull Vet Inst Pulawy37:3–8.
13.
MillerJMOlsonC: 1972, Precipitating antibody to an internal antigen of C-type virus associated with bovine lymphosarcoma. J Natl Cancer Inst49:1459–1461.
14.
MillerJMSchmerrMJF: 1987, Detection of bovine leukemia virus infection by immunodiffusion and radioimmunoassay. In: Enzootic bovine leukosis and bovine leukemia virus, ed. BurnyAMammerickxM, pp. 187–193. Martinus Nijhoff, Boston, MA.
15.
MillerJMVan der MaatenMJ: 1974, A complement-fixation test for bovine leukemia (C-type) virus. J Natl Cancer Inst53:1699–1702.
16.
MillerJMVan der MaatenMJ: 1976, Serologic detection of bovine leukemia virus infection. Vet Microbiol1:195–202.
17.
MurtaughMPLinGFHaggardDL: 1991, Detection of bovine leukemia virus by the polymerase chain reaction. J Virol Methods33:73–85.
18.
NaifHMBrandonRBDanielRCLavinMF: 1990, Bovine leukaemia proviral DNA detection in cattle using the polymerase chain reaction. Vet Microbiol25:117–129.
19.
NaifHMDanielRCCougleWGLavinMF: 1992, Early detection of bovine leukemia virus by using an enzyme-linked assay for polymerase chain reaction-amplified proviral DNA in experimentally infected cattle. J Clin Microbiol30:675–679.
20.
OlsonC: 1974, Bovine lymphosarcoma (leukemia). A synopsis. J Am Vet Med Assoc165:630–632.
21.
PortetelleDBruckCMammerickxMBurnyA: 1983, Use of monoclonal antibody in an ELISA test for the detection of antibodies to bovine leukaemia virus. J Virol Methods6: 1929.
22.
RobertsDHLucasMHWibberleyGChaseyD: 1982, An investigation into susceptibility of cattle to bovine leukosis virus following inoculation by various routes. Vet Rec110:222–224.
23.
SagataNYasunagaTTsuzuku-KawamuraJ: 1985, Complete nucleotide sequence of the genome of bovine leukaemia virus: its evolutionary relationship to other retroviruses. Proc Natl Acad Sci USA82:677–681.
24.
SambrookJFritschEFManiatisT: 1989, Molecular cloning, a laboratory manual, 2nd ed., pp. 9.31–9.56, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
25.
SaundersSBurkeJF: 1990, Isolation of miniprep DNA for double strand sequencing. Nucleic Acids Res18:4948–4952.
26.
ShermanMPErlichGDFerrerJF: 1992, Amplification and analysis of specific DNA and RNA sequences of bovine leukemia virus from infected cows by polymerase chain reaction. J Clin Microbiol30:185–191.
27.
ShettigaraPTSamaghBSLobinowichEM: 1986, Eradication of bovine leukemia virus infection in commercial dairy herds using the agar gel immunodiffusion test. Can J Vet Res50:221–226.
28.
StarcichBRHahnBHShawGM: 1986, Identification and characterization of conserved and variable regions in the envelope gene of HTLV-III/LAV, the retrovirus of AIDS. Cell45:637–648.
29.
VarmaVASwanD: 1990, Polymerase chain reaction: a powerful diagnostic tool. In: Modem pathology of AIDS and other retroviral infections, ed. RaczPHaaseATGluckmanJC, pp. 130–140. Karger, Basel, Switzerland.
30.
WeissRTeichN: 1985, RNA tumor viruses 2, supplements and appendices, 2nd ed., pp. 1089–1101. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
