Quantitation of HTLV-I Proviral Load Using Real-time Quantitative PCR with Taqman MGB Probe
Author(s): XIE Jinzhen, CHEN Changrong, ZHANG Jun, NI Hongying, GE Shengxiang, ZHOU Juanjuan, OU Shanhai, ZHENG Xiujuan, RAN Peng, PEI Bin
Pages: 339-343
Year: 2009 Issue: 5
Journal: Chinese Journal of Virology
Keyword: HTLV-I proviral; Taqman MGB probe; real-time quantitative PCR;
Abstract: 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 β-actin gene.The amplification system was sensitive to detect 5copy/μL.The standard curve had a good linearity when the quantity for the gene was between 103 and 107 copy/μL(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.
Pages: 339-343
Year: 2009 Issue: 5
Journal: Chinese Journal of Virology
Keyword: HTLV-I proviral; Taqman MGB probe; real-time quantitative PCR;
Abstract: 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 β-actin gene.The amplification system was sensitive to detect 5copy/μL.The standard curve had a good linearity when the quantity for the gene was between 103 and 107 copy/μL(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.
Related Articles