Quantitation of HTLV-I proviral load by a TaqMan real-time PCR assay.
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.
Dehée
Axelle
A
Laboratoire de Microbiologie, Hôpital Rothschild, Paris, France.
Césaire
Raymond
R
Désiré
Nathalie
N
Lézin
Agnès
A
Bourdonné
Olivier
O
Béra
Odile
O
Plumelle
Yves
Y
Smadja
Didier
D
Nicolas
Jean Claude
JC
D001483:Base Sequence; D002353:Carrier State; D004279:DNA, Viral; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000254:growth & development; D006801:Humans; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000097:blood / Q000821:virology*; D007963:Leukocytes, Mononuclear / Q000821:virology; D008969:Molecular Sequence Data; D015493:Paraparesis, Tropical Spastic / Q000097:blood / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011533:Proviruses / Q000235:genetics / Q000254:growth & development*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019914:Taq Polymerase; D019562:Viral Load
D004279:DNA, Viral; D019914:Taq Polymerase