BLV-CoCoMo-qPCR-2: improvements to the BLV-CoCoMo-qPCR assay for bovine leukemia virus by reducing primer degeneracy and constructing an optimal standard curve.
Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis, which is the most common neoplastic disease of cattle. Because BLV infection can remain clinically silent, the proviral load is an important index for estimating disease progression. CoCoMo-qPCR, an assay developed to estimate BLV proviral load, allows the highly sensitive detection of BLV originating in different countries. Here, we developed a modified version of the CoCoMo-qPCR assay, the "BLV-CoCoMo-qPCR-2" assay, which uses optimized degenerate primers. We also constructed a new plasmid standard. Finally, we used both assays to examine DNA samples from BLV-infected cattle and compared the results.
Viral Infectious Diseases Unit, RIKEN, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
Shin-nosuke
SN
Takeshima
Yuri
Y
Kitamura-Muramatsu
Yuan
Y
Yuan
Meripet
M
Polat
Susumu
S
Saito
Yoko
Y
Aida
D000818:Animals; D002417:Cattle; D016583:Enzootic Bovine Leukosis / Q000175:diagnosis* / Q000821:virology; D001909:Leukemia Virus, Bovine / Q000235:genetics* / Q000302:isolation & purification*; D025202:Molecular Diagnostic Techniques / Q000379:methods* / Q000592:standards; D015345:Oligonucleotide Probes / Q000235:genetics*; D010957:Plasmids; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D014730:Veterinary Medicine / Q000379:methods / Q000592:standards; D019562:Viral Load / Q000379:methods* / Q000592:standards
D015345:Oligonucleotide Probes