Parekh B, Phillips S, Granade T C, Baggs J, Hu D J, Respess R
Division of HIV, STD, TB Laboratory Research, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
AIDS Res Hum Retroviruses. 1999 Jan 20;15(2):133-42. doi: 10.1089/088922299311556.
We evaluated the performance of three HIV-1 RNA quantitation methods (Amplicor HIV-1 MONITOR-1.0, NASBA, and Quantiplex HIV RNA 2.0 [branched DNA (bDNA)]) using plasma specimens (N = 60) from individuals from Asia and Africa infected with one of three HIV-1 subtypes (A, Thai B [B'] or E; N = 20 each). Our results demonstrate that of the 20 subtype A specimens, 19 were quantifiable by the bDNA assay compared with 15 by the MONITOR-1.0 and 13 by NASBA. Of those quantifiable, the mean log10 difference was 0.93 between bDNA and MONITOR-1.0 and 0.46 between bDNA and NASBA. For subtype B' specimens, the correlation among methods was better with only 2 specimens missed by NASBA and 3 by the bDNA assay. However the missed specimens had viral burden near the lower limit (1000 copies/ml) for these assays. For the 20 subtype E specimens, MONITOR-1.0 and NASBA quantified RNA in 17 and 14 specimens, respectively, as compared with 19 specimens quantified by the bDNA assay. The correlation among different assays, especially between bDNA/NASBA and MONITOR-1.0/NASBA, was poor, although the mean log10 difference for subtype E specimens was 0.4 between bDNA and MONITOR-1.0 and only 0.08 between bDNA and NASBA. The addition of a new primer set, designed for non-B HIV-1 subtypes, to the existing MONITOR assay (MONITOR-1.0+) resulted in RNA detection in all 60 specimens and significantly improved the efficiency of quantitation for subtypes A and E. Our data indicate that HIV-1 subtype variation can have a major influence on viral load quantitation by different methods. Periodic evaluation and modification of these quantitative methods may be necessary to ensure reliable quantification of divergent viruses.
我们使用来自亚洲和非洲感染三种HIV-1亚型（A、泰国B [B'] 或E；各20例）个体的血浆样本（N = 60），评估了三种HIV-1 RNA定量方法（Amplicor HIV-1 MONITOR-1.0、核酸序列扩增技术（NASBA）和Quantiplex HIV RNA 2.0 [分支DNA（bDNA）]）的性能。我们的结果表明，在20份A亚型样本中，19份可通过bDNA检测法定量，相比之下，MONITOR-1.0法可定量15份，NASBA法可定量13份。在可定量的样本中，bDNA与MONITOR-1.0之间的平均log10差异为0.93，bDNA与NASBA之间的平均log10差异为0.46。对于B'亚型样本，各方法之间的相关性较好，NASBA仅漏检2份样本，bDNA检测法漏检3份样本。然而，漏检样本的病毒载量接近这些检测方法的下限（1000拷贝/毫升）。对于20份E亚型样本，MONITOR-1.0和NASBA分别对17份和14份样本中的RNA进行了定量，相比之下，bDNA检测法定量了19份样本。不同检测方法之间的相关性较差，尤其是bDNA/NASBA与MONITOR-1.0/NASBA之间，尽管E亚型样本中bDNA与MONITOR-1.0之间的平均log10差异为0.4，bDNA与NASBA之间仅为0.08。在现有的MONITOR检测法（MONITOR-1.0+）中添加一组为非B型HIV-1亚型设计的新引物，使得所有60份样本均能检测到RNA，并显著提高了A和E亚型的定量效率。我们的数据表明，HIV-1亚型变异可能对不同方法的病毒载量定量产生重大影响。可能需要定期评估和改进这些定量方法，以确保对不同病毒进行可靠的定量。
