Bertine Mélanie, Gueudin Marie, Mélard Adeline, Damond Florence, Descamps Diane, Matheron Sophie, Collin Fidéline, Rouzioux Christine, Plantier Jean-Christophe, Avettand-Fenoel Véronique
IAME, UMR 1137, INSERM, Université Paris Diderot, Sorbonne Paris Cité, Laboratoire de Virologie, Hôpital Bichat, AP-HP, Paris, France.
Laboratoire de Virologie, Hôpital Charles Nicolle, CHU de Rouen, and GRAM, Equipe d'Accueil 2656, Faculté de Médecine-Pharmacie, Institut de Recherche et d'Innovation en Biomédecine, Université de Rouen, Rouen, France.
J Clin Microbiol. 2017 Sep;55(9):2850-2857. doi: 10.1128/JCM.00755-17. Epub 2017 Jul 12.
HIV-2 infection is characterized by a very low replication rate in most cases and low progression. This necessitates an approach to patient monitoring that differs from that for HIV-1 infection. Here, a new highly specific and sensitive method for HIV-2 DNA quantification was developed. The new test is based on quantitative real-time PCR targeting the long terminal repeat (LTR) and regions and using an internal control. Analytical performance was determined in three laboratories, and clinical performance was determined on blood samples from 63 patients infected with HIV-2 group A ( = 35) or group B ( = 28). The specificity was 100%. The 95% limit of detection was three copies/PCR and the limit of quantification was six copies/PCR. The within-run coefficients of variation were between 1.03% at 3.78 log copies/PCR and 27.02% at 0.78 log copies/PCR. The between-run coefficient of variation was 5.10%. Both manual and automated nucleic acid extraction methods were validated. HIV-2 DNA loads were detectable in blood cells from all 63 patients. When HIV-2 DNA was quantifiable, median loads were significantly higher in antiretroviral-treated than in naive patients and were similar for groups A and B. HIV-2 DNA load was correlated with HIV-2 RNA load ( = 0.68; 95% confidence interval [CI], 0.4 to 0.8; < 0.0001). Our data show that this new assay is highly sensitive and quantifies the two main HIV-2 groups, making it useful for the diagnosis of HIV-2 infection and for pathogenesis studies on HIV-2 reservoirs.
在大多数情况下，HIV-2感染的特点是复制率极低且病程进展缓慢。这就需要一种与HIV-1感染患者监测方法不同的方式。在此，开发了一种用于HIV-2 DNA定量的新型高特异性和高灵敏度方法。该新检测方法基于针对长末端重复序列（LTR）和相关区域的定量实时PCR，并使用内部对照。在三个实验室测定了分析性能，并对63例感染HIV-2 A组（n = 35）或B组（n = 28）患者的血样测定了临床性能。特异性为100%。95%检测限为3拷贝/PCR，定量限为6拷贝/PCR。批内变异系数在3.78 log拷贝/PCR时为1.03%，在0.78 log拷贝/PCR时为27.02%。批间变异系数为5.10%。手动和自动核酸提取方法均得到验证。在所有63例患者的血细胞中均可检测到HIV-2 DNA载量。当HIV-2 DNA可定量时，接受抗逆转录病毒治疗患者的中位载量显著高于初治患者，且A组和B组相似。HIV-2 DNA载量与HIV-2 RNA载量相关（r = 0.68；95%置信区间[CI]，0.4至0.8；P < 0.0001）。我们的数据表明，这种新检测方法高度灵敏，可对两个主要HIV-2组进行定量，对HIV-2感染的诊断和HIV-2储存库的发病机制研究具有重要意义。
