[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 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.
Tamegão-Lopes
Bruna Pedroso
BP
Divisão de Biologia Celular e Molecular, Fundação HEMOPA, Belém, PA, Brazil.
Rezende
Priscila Rocha
PR
Maradei-Pereira
Luciana Maria Cunha
LM
de Lemos
José Alexandre Rodrigues
JA
D000328:Adult; D001782:Blood Donors; D004279:DNA, Viral / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015490:HTLV-I Infections / Q000276:immunology / Q000821:virology; D015491:HTLV-II Infections / Q000276:immunology / Q000821:virology; D015368:Human T-lymphotropic virus 1 / Q000235:genetics* / Q000276:immunology; D015367:Human T-lymphotropic virus 2 / Q000235:genetics* / Q000276:immunology; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D011533:Proviruses / Q000235:genetics* / Q000276:immunology; D015203:Reproducibility of Results; D019562:Viral Load / Q000379:methods*
D004279:DNA, Viral