Institute of Tropical Medicine Antwerp
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Development and validation of a multiplex real-time PCR assay for simultaneous genotyping and human T-lymphotropic virus type 1, 2, and 3 proviral load determination

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dc.contributor.author Moens, B.
dc.contributor.author López, G.
dc.contributor.author Adaui, V.
dc.contributor.author González, E.
dc.contributor.author Kerremans, L.
dc.contributor.author Clark, D.
dc.contributor.author Verdonck, K.
dc.contributor.author Gotuzzo, E.
dc.contributor.author Vanham, G.
dc.contributor.author Cassar, O.
dc.contributor.author Gessain, A.
dc.contributor.author Vandamme, A. M.
dc.contributor.author Van Dooren, S.
dc.date.accessioned 2009-12-21T15:22:22Z
dc.date.available 2009-12-21T15:22:22Z
dc.date.issued 2009
dc.identifier.issn 0095-1137
dc.identifier.doi http://dx.doi.org/10.1128/JCM.00781-09
dc.identifier.other ITG-M7B
dc.identifier.other ITG-M9A
dc.identifier.other MICRO
dc.identifier.other U-VIROL
dc.identifier.other JIF
dc.identifier.other DOI
dc.identifier.other UPD17
dc.identifier.other ABSTRACT
dc.identifier.uri http://hdl.handle.net/10390/2870
dc.description.abstract The human T-lymphotropic virus (HTLV) proviral load remains the best surrogate marker for disease progression. Real-time PCR techniques have been developed for detection and quantification of cosmopolitan HTLV type 1a (HTLV-1a) and HTLV-2. Since a growing level of diversity in subtypes and genotypes is observed, we developed a multiplex quantitative PCR for simultaneous detection, genotyping, and quantification of proviral loads of HTLV-1, 2, and 3. Our assay uses tax type-specific primers and dually labeled probes and has a dynamic range of 10(5) to 10 HTLV copies. One hundred sixty-three samples were analyzed, among which all of the different subtypes within each HTLV genotype could be detected. The performance of proviral load determination of our multiplex assay was compared with that of a previously published HTLV-1 singleplex quantitative PCR based on SYBR green detection, developed at a different institute. Linear regression analysis showed a statistically significant (P < 0.0001) and strong (r(2) = 0.87) correlation between proviral load values measured with the two distinct real-time PCR assays. In conclusion, our novel assay offers an accurate molecular diagnosis and genotyping, together with the determination of the proviral load of HTLV-infected individuals, in a single amplification reaction. Moreover, our molecular assay could offer an alternative when current available serological assays are insufficient. en
dc.language English en
dc.subject Viral diseases en
dc.subject HTLV en
dc.subject Human T-lymphotropic virus en
dc.subject Genotyping en
dc.subject Proviral load en
dc.subject Assays en
dc.subject Multiplex PCR en
dc.subject Real-time en
dc.title Development and validation of a multiplex real-time PCR assay for simultaneous genotyping and human T-lymphotropic virus type 1, 2, and 3 proviral load determination en
dc.type Article en
dc.citation.issue 11 en
dc.citation.jtitle Journal of Clinical Microbiology en
dc.citation.volume 47 en
dc.citation.pages 3682-3691 en
dc.identifier.pmid http://www.ncbi.nlm.nih.gov/pubmed/19741085
dc.citation.jabbreviation J Clin Microbiol en


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