Resumo
The present study aimed at investigating the best algorithm definition to be employed for diagnosing the human T lymphotropic virus type 1 (HTLV-1) and type 2 (HTLV-2) in HIV-1 infected/Aids patients. Blood samples of 1,608 HIV-1 infected patients from the AIDS Reference Center (CRT DST/AIDS-SP) were tested for the presence of HTLV-1/2 antibodies using two screening assays (EIA Murex HTLV-I+II, and Gold ELISA HTLV-I/II), and confirmed by two Blots [HTLV Blot 2.4 (Western Blot WB) and INNO-LIA HTLV I/II (Line ImmunoAssay - LIA)], and one molecular assay (pol real-time PCR). The screening tests detected 51(Murex) and 49 (Gold ELISA) reagents sera. WB , confirmed 23 HTLV-1, 12 HTLV-2, six HTLV, and nine showed HTLV indeterminate profiles. LIA confirmed 24 HTLV-1, 20 HTLV-2, and six HTLV. PCR detected 18 HTLV-1- and 12 HTLV-2-infected blood samples. By using any confirmatory assay, 50 patients confirmed HTLV infection: 25 HTLV-1 (1.55 %), 21 HTLV-2 (1.31 %) and four HTLV (0.25 %). The sensitivity of LIA, WB and PCR assays were 96 %, 76 % and 60 %, respectively. By considering the assays cost as the sole variable, the best testing algorithm for diagnosing HIV-1/HTLV-coinfection in HIV-1 infected patient was the use of PCR followed by LIA technique(AU)
O presente estudo pesquisou o melhor algoritmo de testes laboratoriais para efetuar o diagnóstico de infecção por vírus linfotrópicos de células T humanas dos tipos 1 (HTLV-1) e 2 (HTLV-2) em pacientes HIV-1 positivos. Amostras de sangue de 1.608 pacientes do CRT DST/Aids-SP foram analisadas quanto à presença de anticorpos específicos usando-se dois ensaios de triagem (EIA Murex HTLV-I+II e Gold ELISA HTLV-I/II), dois confirmatórios [HTLV Blot 2.4 (Western Blot WB) e INNO-LIA HTLV I/II (Line ImmunoAssay - LIA)] e um molecular (PCR em tempo real pol). Na triagem foram detectados 51(Murex) e 49 (Gold ELISA) soros reagentes. Pelo WB, 23 soros confirmaram infecção por HTLV-1, 12 HTLV-2, seis HTLV e nove apresentaram perfis indeterminados. O LIA detectou 24 soros HTLV-1 positivos, 20 HTLV-2 e seis HTLV. A PCR evidenciou segmento pol de HTLV-1 em 18 e HTLV-2 em 12 amostras de sangue. Pelos testes confirmatórios, em 50 pacientes foi confirmada a infecção por HTLV: 25 HTLV-1 (1,55 %), 21 HTLV-2 (1,31 %) e quatro HTLV (0,25 %). As sensibilidades do LIA, WB e PCR foram de 96 %, 76 % e 60 %, respectivamente. Considerando-se apenas o custo, o melhor algoritmo diagnóstico para população infectada pelo HIV-1 foi o uso da PCR seguida do LIA(AU)
Assuntos
Humanos , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Vírus Linfotrópico T Tipo 2 Humano/isolamento & purificação , Testes Laboratoriais/análise , Testes Laboratoriais/métodos , HIV-1 , Testes Sorológicos/métodos , Reação em Cadeia da Polimerase em Tempo Real , Coinfecção/diagnósticoResumo
A cross-sectional study was performed on HIV-1 infected individuals with or without antiretroviral treatment (ARV) in the AIDS Day Hospital, Botucatu Medical School, UNESP. Between August 2004 and October 2005, 73 HIV-1 infected individuals were divided into three groups: infected individuals with or without AIDS who had never received ARV (G1 = 15); patients on HAART that had had plasma HIV-1 RNA viral load (VL) equal to or greater than 50 copies/mL (G2 = 27); and patients on HAART with undetectable VL for at least the past six months (G3 = 31). There was also an additional group that comprised blood donors without any sign of the disease and with negative HIV serum tests (G4 = 20), which was the control group. Serum cytokine levels (values in pg/mL) were measured by enzyme-linked immunosorbent assay (ELISA) and specific mRNA expression by reverse transcription polymerase chain reaction (RT-PCR). Both techniques were performed on the four groups for TNF-á, IL-2, INF-ã, IL-4 and IL-10. All patients were submitted to VL determination and CD4+ and CD8+T lymphocyte counts. The analysis of the results revealed a significant comparison among groups for both methods and an association between the latter (> 80% r² > 0.80). There was only one exception, in control individuals for IL-2 by ELISA. The cytokine profiles, in both methods, for the three patient groups, were mature Th-0. The behaviors of IL-2 and INF-ã required emphasis due to consequent expression of dominant Th profile. Both methods showed low IL-2 and high mean values of INF-ã in the three groups. Several authors have recently drawn attention to the substantial apoptosis of infected and non-infected CD4+T cells, mainly during primary infection, persisting only in those with INF-ã phenotype producer and not IL-2. HIV infected individuals submitted to HAART are expected to produce IL-2 in an attempt to present Th-1 profile, but in most cases this did not occur.(AU)