Transfer and evaluation of an automated, low-cost real-time reverse transcription-PCR test for diagnosis and monitoring of human immunodeficiency virus type 1 infection in a west African resource-limited setting
Transfer and evaluation of an automated, low-cost real-time reverse transcription-PCR test for diagnosis and monitoring of human immunodeficiency virus type 1 infection in a west African resource-limited setting
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DOI：
10.1128/jcm.43.6.2709-2717.2005
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发表时间：
2005-06-01
影响因子：
9.4
通讯作者：
Rouzioux, C
中科院分区：
文献类型：
--
作者：
Rouet, F;Ekouevi, DK;Rouzioux, C
在资源有限的环境中迫切需要低成本的人类免疫缺陷病毒 1 型 (HIV-1) 病毒载量 (VL) 监测技术。自动 TaqMan 实时逆转录 PCR (RT-PCR) 检测被转移到科特迪瓦阿比让 SIDA 诊断和研究中心的实验室，并对四个 ANRS 研究项目收集的 806 份血浆样本进行 HIV-1 RNA VL 检测评估。首先确定测定的检测阈值和再现性。在主要含有循环重组型 02 毒株 (CRF02) 的样本中，将通过该测定获得的定量结果与两种商业 HIV-1 RNA 试剂盒（Versant 3.0 版和 Monitor 1.5 版测定）进行比较。该测试的临床评估是在不同情况下进行的，包括儿科感染的早期诊断和抗逆转录病毒治疗患者的监测。我们方法的定量限为 300 拷贝/ml。通过实时 PCR 测定获得的 HIV-1 RNA 值与通过 Versant 试剂盒（r = 0.901；P < 0.001）和 Monitor 测试（r = 0.856；P < 0.001）获得的值高度相关，并且根据 HIV-1 基因型均匀分布。对于儿科 HIV-1 感染的早期诊断，与 Versant 结果相比，实时 PCR 检测的敏感性和特异性均为 100%（95% 置信区间分别为 93.7 至 100.0 和 98.3 至 100.0）。开始抗逆转录病毒治疗后，HIV-1 RNA 水平的动力学非常相似，在使用我们的技术和 Versant 检测进行随访期间，成人和儿童的相似比例低于检测限。 TaqMan 实时 PCR（每次测试 12 美元）现在在我们的实验室中常规用于监测 HIV-1 感染。应在 CRF02 以外的菌株流行的资源有限国家进一步评估这项技术。
There is an urgent need for low-cost human immunodeficiency virus type 1 (HIV-1) viral load (VL) monitoring technologies in resource-limited settings. An automated TaqMan real-time reverse transcription-PCR (RT-PCR) assay was transferred to the laboratory of the Centre de Diagnostic et de Recherches sur le SIDA, Abidjan, Cote d'Ivoire, and assessed for HIV-1 RNA VL testing in 806 plasma samples collected within four ANRS research programs. The detection threshold and reproducibility of the assay were first determined. The quantitative results obtained with this assay were compared with two commercial HIV-1 RNA kits (the Versant version 3.0 and Monitor version 1.5 assays) in specimens harboring mainly the circulating recombinant form 02 strain (CRF02). The clinical evaluation of this test was done in different situations including the early diagnosis of pediatric infection and the monitoring of antiretroviral-treated patients. The quantification limit of our method was 300 copies/ml. The HIV-1 RNA values obtained by real-time PCR assay were highly correlated with those obtained by the Versant kit (r = 0.901; P < 0.001) and the Monitor test (r = 0.856; P < 0.001) and homogeneously distributed according to HIV-1 genotypes. For the early diagnosis of pediatric HIV-1 infection, the sensitivity and specificity of the real-time PCR assay were both 100% (95% confidence intervals of 93.7 to 100.0 and 98.3 to 100.0, respectively), compared to the Versant results. Following initiation of antiretroviral treatment, the kinetics of HIV-1 RNA levels were very comparable, with a similar proportion of adults and children below the detection limit during follow-up with our technique and the Versant assay. The TaqMan real-time PCR ($12 per test) is now routinely used to monitor HIV-1 infection in our laboratory. This technology should be further evaluated in limited-resource countries where strains other than CRF02 are prevalent.