[Quantitation of HTLV-I proviral load using real-time quantitative PCR with Taqman MGB probe].
A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood. The technology utilizes special primers and Taqman MGB fluorescence probe to measure amplification products from the gag-pro-pol polyprotein gene of HTLV-I. HTLV-I copy number was normalized to the amount of cellular DNA by quantitation of the beta-actin gene, The amplification system was sensitive to detect 5 copy/microL. The standard curve had a good linearity when the quantity for the gene was between 10(3) and 10(7) copy/microL (R2 = 0.999). Good reproducibility was observed in each intra- and inter-assay. We also measured proviral load in peripheral blood in 12 HTLV-I seropositive former blood donors. Proviral load for HTLV-I infected donors ranged from 0.015 to 12.819 copy/cell in WBC with the mean of 3.116 copy/cell.
Xie
Jin-Zhen
JZ
Xiamen Blood Services, Xiamen 361004, China.
Chen
Chang-Rong
CR
Zhang
Jun
J
Ni
Hong-Ying
HY
Ge
Sheng-Xiang
SX
Zhou
Juan-Juan
JJ
Ou
Shan-Hai
SH
Zheng
Xiu-Juan
XJ
Ran
Peng
P
Pei
Bin
B
D015683:Gene Products, gag / Q000235:genetics; D015684:Gene Products, pol / Q000235:genetics; D015368:Human T-lymphotropic virus 1 / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D015335:Molecular Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D014764:Viral Proteins / Q000235:genetics
D015683:Gene Products, gag; D015684:Gene Products, pol; D015335:Molecular Probes; D014764:Viral Proteins