Importance of primer selection in the application of PCR technology to the diagnosis of bovine leukemia virus.
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 microliters 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.
Marsolais
G
G
Ministère de l'Agriculture des Pêcheries et de l'Alimentation du Québec, Canada.
Dubuc
R
R
Bergeron
J
J
Morrey
J D
JD
Kelly
E J
EJ
Jackson
M K
MK
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D017931:DNA Primers; D004279:DNA, Viral / Q000302:isolation & purification; D016583:Enzootic Bovine Leukosis / Q000097:blood / Q000821:virology*; D001909:Leukemia Virus, Bovine / Q000302:isolation & purification*; D007962:Leukocytes / Q000821:virology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011792:Quebec; D012680:Sensitivity and Specificity; D014589:Utah
D017931:DNA Primers; D004279:DNA, Viral