Immunology Department, Instituto Adolfo Lutz, Coordenadoria de Controle de Doenças da Secretaria de Estado da Saúde de São Paulo, Av. Dr. Arnaldo, 355, 11° andar, 01246-902, São Paulo, SP, Brazil.
J Virol Methods. 2011 May;173(2):280-6. doi: 10.1016/j.jviromet.2011.02.018. Epub 2011 Feb 22.
The ability to confirm the diagnosis of human T-lymphotropic virus types 1 and 2 (HTLV-1 and HTLV-2) in at-risk individuals in São Paulo, Brazil by Western blotting (WB), conventional polymerase chain reaction (tax and pol PCR) and real-time PCR (pol) is compared. Seventy-three blood samples that were reactive in HTLV-1/2 serological screening enzyme immunoassays (EIAs) were evaluated. HTLV-1/2 was confirmed in 53 blood samples: 48 were positive by WB, 41 were positive by PCR and 42 scored positive by real-time PCR assays (37 of 48 WB-positive samples plus five WB-indeterminate samples that were further confirmed by sequencing). Although WB was able to detect more cases of HTLV-1/2 infection, the real-time PCR assay was able to discriminate between these two viruses and confirm an individual HTLV-1/HTLV-2 diagnosis in two HTLV WB-untyped samples and five WB-indeterminate samples. Because of the large number of WB-indeterminate samples and the cost of the WB assay in Brazil, it is proposed an algorithm that employs two EIAs for screening and then real-time PCR to confirm the infection, followed by testing any PCR-negative samples with the WB assay. This strategy reduces costs and improves the accuracy of the diagnosis of HTLV-1/2.
比较了在巴西圣保罗,通过 Western blot(WB)、常规聚合酶链反应(tax 和 pol PCR)和实时 PCR(pol)来确认人类 T 淋巴细胞病毒 1 型和 2 型(HTLV-1 和 HTLV-2)感染高危个体的诊断能力。评估了 73 份在 HTLV-1/2 血清学筛查酶免疫测定(EIA)中呈反应性的血液样本。在 53 份血液样本中确认了 HTLV-1/2:48 份通过 WB 阳性,41 份通过 PCR 阳性,42 份通过实时 PCR 检测阳性(37 份 WB 阳性样本加 5 份 WB 不确定样本,通过测序进一步确认)。尽管 WB 能够检测到更多的 HTLV-1/2 感染病例,但实时 PCR 检测能够区分这两种病毒,并在 2 份 HTLV WB 未分型样本和 5 份 WB 不确定样本中确认了个体 HTLV-1/HTLV-2 诊断。由于 WB 不确定样本数量较多,以及巴西 WB 检测的成本较高,因此建议采用两种 EIA 进行筛查,然后使用实时 PCR 进行确认感染,然后对任何 PCR 阴性样本进行 WB 检测。这种策略降低了成本并提高了 HTLV-1/2 的诊断准确性。