Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay
Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay
复制标题
DOI：
10.1016/s0166-0934(01)00445-1
复制
发表时间：
2002-04-01
影响因子：
3.1
通讯作者：
Nicolas, JC
中科院分区：
文献类型：
--
作者：
Dehée, A;Césaire, R;Nicolas, JC
采用实时荧光定量PCR方法测定人t淋巴细胞嗜I型病毒（HTLV-I）在外周血单个核细胞（PBMCs）中的前病毒载量。HTLV-I拷贝数是指通过白蛋白基因的定量细胞DNA的实际数量。10份HTLV-I DNA的检测灵敏度为100%，且检测范围至少为5 log，，。通过从htlv -i感染患者中独立提取pbmc来评估检测内和检测间的重复性（变异系数分别为24%和7%）。TaqMan PCR检测的性能，加上其高通量，因此可以对感染患者的HTLV-I前病毒载量进行可靠的常规随访。临床样本的初步结果表明，htlv - 1相关脊髓病/热带痉挛性截瘫患者的前病毒载量高于无症状携带者，这也表明这种定量测量方法在评估htlv - 1与成人t细胞白血病/淋巴瘤样综合征之间的病因学联系方面是有用的。(C) 2002 Elsevier Science B.V.版权所有
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 mononuclear cells (PBMCs). The HTLV-I copy number was referred to the actual amount of cellular DNA by means of the quantitation of the albumin gene. Ten copies of HTLV-I DNA could be detected with 100%,, sensitivity, and the assay had a wide range of at least 5 log,,. Intra- and inter-assay reproducibility was evaluated using independent extractions of PBMCs from an HTLV-I-infected patient (coefficients of variation, 24 and 7% respectively). The performance of this TaqMan PCR assay, coupled with its high throughput, thus allows reliable routine follow-up of HTLV-I proviral load in infected patients. Preliminary results using clinical samples indicate a higher proviral load in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis than in asymptomatic carriers, and also suggest the usefulness of this quantitative measurement to assess the etiological link between HTLV-I and adult T-cell leukaemia/lymphoma-like syndromes. (C) 2002 Elsevier Science B.V. All rights reserved.