Avettand-Fenoel Véronique, Damond Florence, Gueudin Marie, Matheron Sophie, Mélard Adeline, Collin Gilles, Descamps Diane, Chaix Marie-Laure, Rouzioux Christine, Plantier Jean-Christophe
Laboratoire de Virologie, AP-HP, Hôpital Necker Enfants Malades, Paris, France EA7327, Université Paris-Descartes, Sorbonne Paris Cité, Faculté de Médecine, Paris, France.
INSERM, IAME, UMR 1137, Paris, France Université Paris Diderot, Sorbonne Paris Cité, Paris, France AP-HP, Hôpital Bichat-Claude Bernard, Laboratoire de Virologie, Paris, France.
J Clin Microbiol. 2014 Aug;52(8):3017-22. doi: 10.1128/JCM.00724-14. Epub 2014 Jun 11.
The Agence Nationale de Recherche sur le Sida et les hépatites virales (ANRS) previously developed a widely used method for HIV-1 RNA quantification (Biocentric). Here, we report the development of a new specific and sensitive method for HIV-2 RNA quantification, based on an adaptation of the existing HIV-1 protocol. The new test is based on TaqMan one-step reverse transcription-quantitative PCR (qRT-PCR) targeting two conserved consensus regions of HIV-2 (long terminal repeat [LTR] and gag). Analytic performances were determined in three laboratories. Clinical performances were evaluated on 100 plasma samples from HIV-2-infected patients (groups A, B, and H) by comparison with the assay currently used for the ANRS HIV-2 cohort. The specificity was 100%. Sensitivity was 50 copies/ml (cp/ml) and was optimized to 10 cp/ml. The within-run coefficients of variation in the three laboratories varied from 0.54% to 1.61% at 4 log10 copies/ml and from 7.24% to 14.32% at 2 log10 cp/ml. The between-run coefficients of variation varied from 2.28% to 6.43%. Of the 39 clinical samples below 2 log10 in the current assay, the new test improved the detection or quantification of 17 samples, including eight group B samples. For quantifiable samples, similar loads were obtained with the two assays for group A samples. The median difference between the two assays for group B samples was +0.18 but with greater heterogeneity than for group A. The HIV-2 group H sample had similar results with the two assays. This new assay is highly sensitive and accurately quantifies the most prevalent HIV-2 groups. This test will be useful for monitoring low viral loads in HIV-2-infected patients.
法国国家艾滋病和病毒性肝炎研究机构（ANRS）此前开发了一种广泛使用的HIV-1 RNA定量方法（Biocentric）。在此，我们报告基于对现有HIV-1方案的改进，开发了一种用于HIV-2 RNA定量的新型特异性和灵敏性方法。新检测方法基于TaqMan一步法逆转录定量PCR（qRT-PCR），靶向HIV-2的两个保守共有区域（长末端重复序列[LTR]和gag）。在三个实验室中确定了分析性能。通过与目前用于ANRS HIV-2队列的检测方法进行比较，对100份来自HIV-2感染患者（A组、B组和H组）的血浆样本进行了临床性能评估。特异性为100%。灵敏度为50拷贝/毫升（cp/ml），并优化至10 cp/ml。三个实验室的批内变异系数在4 log10拷贝/ml时为0.54%至1.61%，在2 log10 cp/ml时为7.24%至14.32%。批间变异系数为2.28%至6.43%。在当前检测中低于2 log10的39份临床样本中，新检测方法改进了17份样本的检测或定量，包括8份B组样本。对于可定量样本，A组样本的两种检测方法获得了相似的载量。B组样本两种检测方法之间的中位数差异为+0.18，但异质性高于A组。HIV-2 H组样本的两种检测方法结果相似。这种新检测方法高度灵敏，能准确量化最常见的HIV-2组。该检测方法将有助于监测HIV-2感染患者的低病毒载量。
