1. Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay.
- Author
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Dehée A, Césaire R, Désiré N, Lézin A, Bourdonné O, Béra O, Plumelle Y, Smadja D, and Nicolas JC
- Subjects
- Base Sequence, Carrier State, DNA, Viral, Human T-lymphotropic virus 1 genetics, Human T-lymphotropic virus 1 growth & development, Humans, Leukemia-Lymphoma, Adult T-Cell blood, Leukocytes, Mononuclear virology, Molecular Sequence Data, Paraparesis, Tropical Spastic blood, Proviruses genetics, Reproducibility of Results, Sensitivity and Specificity, Taq Polymerase, Leukemia-Lymphoma, Adult T-Cell virology, Paraparesis, Tropical Spastic virology, Polymerase Chain Reaction methods, Proviruses growth & development, Viral Load
- 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 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(10). 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.
- Published
- 2002
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