HTLV-1 and HTLV-2 proviral load: a simple method using quantitative real-time PCR

When the human T cell lymphotropic virus (HTLV) is integrated with the host cell genome (provirus), its proviral DNA is a replication marker. Proviral load appears to be an important factor in the development of diseases related to these retroviruses. In this study, a methodology for absolute quanti...

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Bibliographic Details
Published in:Revista da Sociedade Brasileira de Medicina Tropical 2006-11, Vol.39 (6), p.548
Main Authors: Tamegão-Lopes, Bruna Pedroso, Rezende, Priscila Rocha, Maradei-Pereira, Luciana Maria Cunha, de Lemos, José Alexandre Rodrigues
Format: Article
Language:Portuguese
Subjects:
Adult
Blood Donors
DNA, Viral - analysis
Enzyme-Linked Immunosorbent Assay
Female
HTLV-I Infections - immunology
HTLV-I Infections - virology
HTLV-II Infections - immunology
HTLV-II Infections - virology
Human T-lymphotropic virus 1 - genetics
Human T-lymphotropic virus 1 - immunology
Human T-lymphotropic virus 2 - genetics
Human T-lymphotropic virus 2 - immunology
Humans
Male
Middle Aged
Polymerase Chain Reaction - methods
Proviruses - genetics
Proviruses - immunology
Reproducibility of Results
Viral Load - methods
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Summary:When the human T cell lymphotropic virus (HTLV) is integrated with the host cell genome (provirus), its proviral DNA is a replication marker. Proviral load appears to be an important factor in the development of diseases related to these retroviruses. In this study, a methodology for absolute quantification of the HTLV-1 and HTLV-2 proviral load using real-time PCR was developed. Fifty-three blood donor samples with positive ELISA test result were subjected to this methodology, which utilized the TaqMan system for three target sequences: HTLV-1, HTLV-2 and albumin. The absolute proviral load was quantified using the relative ratio between the HTLV genome and the host cell genome, taking into consideration the white blood cell count. The method presented is sensitive (215 copies/ml), practical and simple for proviral quantification, and is efficient and appropriate for confirming and discriminating infections according to viral type.
ISSN:0037-8682
1678-9849
DOI:10.1590/S0037-86822006000600007