[PCR-based detection of proviral DNA of bovine immunodeficiency virus].
A set of primers was developed to detect by polymerase chain reaction (PCR) the proviral DNA of bovine immunodeficiency virus (BIV). A short fragment of 101 bp BIV gene was selected as a target for primers; sequences of proviral DNA isolated from both a cell culture with BIV and from lymphocytes of an experimentally infected animal were known for the fragment. An amplicon of an expected size was detected by standard PCR in a transformed cell series of bovine testicles with Florida 112 BIV DNA, and in a plasmid DNA with a cloned proviral DNA of R29 BIV. Described in the paper are the results of a theoretical comparison of primers used in the detection of BIV by PCR. The presence of non-complementary nucleotides in the set of "primer-single stranded amplicon" was shown to bring about false positive results in the detection of BIV by PCR. No 1500 bp PCR product was detected after PCR with a synthesized pair of primers and with 100% homology for all known BIV isolates complementary to env gene. Finally, the issue of how to detectVIR in clinical samples obtained from experimentally and naturally infected is discussed.
Limanskaia
O
O
Rola
M
M
Bicka
L
L
Kuzmak
J
J
Limanskiĭ
A
A
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000097:blood / Q000821:virology; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D015686:Gene Products, env / Q000235:genetics; D016087:Immunodeficiency Virus, Bovine / Q000235:genetics / Q000302:isolation & purification*; D016180:Lentivirus Infections / Q000662:veterinary / Q000821:virology; D008214:Lymphocytes / Q000821:virology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification*; D016415:Sequence Alignment
D017931:DNA Primers; D004279:DNA, Viral; D015686:Gene Products, env