Ultrasensitive monitoring of HIV-I viral load by a low-cost real-time reverse transcription-PCR assay with internal control for the 5′ long terminal repeat domain
Ultrasensitive monitoring of HIV-I viral load by a low-cost real-time reverse transcription-PCR assay with internal control for the 5′ long terminal repeat domain
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DOI：
10.1373/clinchem.2006.066498
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发表时间：
2006-07-01
影响因子：
9.3
通讯作者：
Preiser, Wolfgang
中科院分区：
文献类型：
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作者：
Drosten, Christian;Panning, Marcus;Preiser, Wolfgang
背景：目前的HIV-1病毒载量测定对于资源有限的环境来说过于昂贵。在一些国家，监测抗逆转录病毒治疗现在比治疗本身更昂贵。此外，一些商业分析在量化罕见基因型方面存在缺陷。方法：我们评估了实时逆转录pcr与内部控制针对HIV-1保守的长末端重复（LTR）结构域的参考面板。以及来自巴西（n = 1186）、南非（n = 130）、印度（n = 44）和德国（n = 127）的患者样本。结果：血浆HIV-1 RNA检出限为31.9 IU /mL （Probit分析，检出概率为95%）。内对照对3.7%的样品有抑制作用（95%置信区间2.32% ~ 5.9%;n = 454； 40个不同的运行）。比较定性检测结果如下：罗氏Amplicor vs LTR检测（n = 431个样本），51.7% vs 65%阳性；Amplicor超敏vs LTR (n = 133), 81.2% vs 82.7%；BioMerieux NucliSens HIV-1 QT (n = 453), 60.5% vs 65.1%；拜耳Versant 3.0 (n = 433), 57.7% vs 55.4%a；总数（n = 1450）， 59.0% vs 63.8%阳性。在中浓度和接近阴性浓度下，测定内/测定间变异率为18%-51%。定量范围50 ~ 10 000 000 IU/mL。与Amplicor相比，A-D、F-J、AE和AG亚型的病毒载量在103-104 IU/mL范围内的平均差异为0.31 log（10）。Amplicor未检测到HIV-1 N和O，但在LTR试验中产量高达180 180.00 IU/mL。与Amplicor、NucliSens或Versant相比，所有国家储存样本中的病毒载量产生的回归线斜率（SD）为0.9(0.13)（所有的P < 0.001）。结论：该方法具有商业检测的所有特点，涵盖所有相关基因型。它可以在资源有限的情况下对抗逆转录病毒治疗进行全面监测。(c) 2006美国临床化学协会。
Background: Current HIV-1 viral-load assays are too expensive for resource-limited settings. In some countries, monitoring of antiretroviral therapy is now more expensive than treatment itself. In addition, some commercial assays have shown shortcomings in quantifying rare genotypes.Methods: We evaluated real-time reverse transcription-PCR with internal control targeting the-conserved long terminal repeat (LTR) domain of HIV-1 on reference panels. and patient samples from Brazil (n = 1186), South Africa (n = 130), India (n = 44), and Germany (n = 127).Results: The detection limit was 31.9 IU of HIV-1 RNA/mL of plasma (> 95% probability of detection, Probit analysis). The internal control showed inhibition in 3.7% of samples (95% confidence interval, 2.32%-5.9%; n = 454; 40 different runs). Comparative qualitative testing yielded the following: Roche Amplicor vs LTR assay (n = 431 samples), 51.7% vs 65% positives; Amplicor Ultrasensitive vs LTR (n = 133), 81.2% vs 82.7%; BioMerieux NucliSens HIV-1 QT (n = 453), 60.5% vs 65.1%; Bayer Versant 3.0 (n = 433), 57.7% vs 55.4%a; total (n = 1450), 59.0% vs 63.8% positives. Intra-/interassay variability at medium and near-negative concentrations was 18%-51%. The quantification range was 50-10 000 000 IU/mL. Viral loads for subtypes A-D, F-J, AE, and AG yielded mean differences of 0.31 log(10) compared with Amplicor in the 103-104 IU/mL range. HIV-1 N and O were not detected by Amplicor, but yielded-up to 180 180.00 IU/mL in the LTR assay. Viral loads in stored samples from all countries, compared with Amplicor, NucliSens, or Versant, yielded regression line slopes (SD) of 0.9 (0.13) (P < 0.001 for all).Conclusions: This method offers all features of commercial assays and covers all relevant genotypes. It could allow general monitoring of antiretroviral therapy in resource-limited settings. (c) 2006 American Association for Clinical Chemistry.