Institute of Virology Helmut-Ruska-Haus, Charité University Medicine Berlin, Berlin, Germany.
Transfusion. 2010 Mar;50(3):685-92. doi: 10.1111/j.1537-2995.2009.02477.x. Epub 2009 Nov 9.
Human immunodeficiency virus Type 1 (HIV-1) assays applying nucleic acid testing (NAT) rely on HIV-1 sequence-specific primers and probes. Their hybridization can be limited or abolished by genetic polymorphisms occurring in the target sequence.
Blood donations are routinely tested for HIV-1/2 antibodies and for HIV-1 RNA in our blood transfusion unit. Recently, HIV-1 RNA was undetectable with an established in-house real-time long terminal repeat (LTR) reverse transcriptase-polymerase chain reaction (RT-PCR) in two cases, whereas serologic assays were positive. The reason for this discrepancy was elucidated by sequencing of the NAT target region in the respective single donations. An improved primer was designed and tested on HIV-1 reference panels and blood donations to ensure reliable detection of HIV-1 RNA.
Direct sequencing of the target region, isolated from samples of two unrelated HIV-positive blood donors, revealed one and four mismatches in the hybridization domain of the forward primer, respectively. Both viruses belong to HIV-1 Subtype B. LTR RT-PCR with an additional forward primer was suitable for all strains of HIV-1 tested with high sensitivity.
Surveillance of HIV-1 genetic diversity is essentially required to continually evaluate its impact on performance of diagnostic and patient monitoring assays.
应用核酸检测（NAT）的人类免疫缺陷病毒 1 型（HIV-1）检测方法依赖于 HIV-1 序列特异性引物和探针。在目标序列中发生的遗传多态性可能会限制或破坏它们的杂交。
在我们的输血单位，血液捐献者通常会接受 HIV-1/2 抗体和 HIV-1 RNA 的检测。最近，在两种情况下，经过建立的内部实时长末端重复（LTR）逆转录酶-聚合酶链反应（RT-PCR）检测 HIV-1 RNA 不可检测，而血清学检测呈阳性。这种差异的原因是通过对各自的单个供体的 NAT 目标区域进行测序来阐明的。设计了一种改进的引物，并在 HIV-1 参考面板和血液捐献物上进行了测试，以确保可靠检测 HIV-1 RNA。
直接对来自两个不相关的 HIV 阳性血液供体样本的目标区域进行测序，分别在正向引物的杂交区域发现了一个和四个错配。两种病毒均属于 HIV-1 亚型 B。带有额外正向引物的 LTR RT-PCR 适用于所有测试的 HIV-1 株，具有很高的灵敏度。
持续评估 HIV-1 遗传多样性对诊断和患者监测检测的性能的影响，因此对 HIV-1 遗传多样性的监测是必不可少的。
