Manigart Olivier, Courgnaud Valérie, Sanou Oumar, Valéa Diane, Nagot Nicolas, Meda Nicolas, Delaporte Eric, Peeters Martine, Van de Perre Philippe
Centre Muraz, 01 BP 390 Bobo-Dioulasso, Burkina Faso.
AIDS. 2004 Aug 20;18(12):1645-51. doi: 10.1097/01.aids.0000131333.30548.db.
To describe and evaluate a simple procedure to identify HIV-1 co- or super-infections based on a heteroduplex mobility assay (HMA).
To identify heteroduplexes corresponding to divergent viral populations in a the same individual, HMA was applied to single DNA samples from each subject in a prospective cohort of commercial sex workers (CSW) in Bobo-Dioulasso, Burkina Faso. After denaturation and renaturation of env DNA amplicons, hybridized DNA was separated by electrophoresis through polyacrylamide gel. HIV-1 co-infections were suspected by slow migration of heteroduplex, at a level comparable to that of mixed reference strains. Following electrophoresis, DNA in bands representing heteroduplex was extracted and cloned in a plasmid vector; identification of phylogenetically distinct clones was confirmed by sequencing divergent clones identified through a second HMA step of a pair of two mixed clones.
Among 147 HIV-1 infected CSW, four had an autologous HMA profile comparable to low mobility of hybridized DNA from mixed reference strains representing most frequent HIV-1 clades and circulating recombinant forms (CRF) prevalent in Burkina Faso. In two of them, two phylogenetically distinct HIV-1 populations were coexisting. The first woman presented with a CRF02-AG and CRF06-cpx co-infection, and the second with a CRF02-AG and a divergent virus co-infection not significantly related to any other known subtypes. In both women, retrospective analyses of stored samples by the same HMA procedure showed acquisition of a second virus concomitent with an increasing plasma HIV RNA.
Autologous HMA procedure followed by acrylamide extraction of heteroduplexes allowed identifying HIV-1 co- and super-infections in a large cohort study. HIV-1 super-infection is not an uncommon phenomenon.
描述并评估一种基于异源双链迁移分析（HMA）来鉴定HIV-1合并感染或重叠感染的简单方法。
为了鉴定同一受试者中与不同病毒群体相对应的异源双链，在布基纳法索博博迪乌拉索的一个商业性工作者（CSW）前瞻性队列中，将HMA应用于每个受试者的单个DNA样本。在env DNA扩增子变性和复性后，通过聚丙烯酰胺凝胶电泳分离杂交的DNA。异源双链迁移缓慢，其水平与混合参考菌株相当，则怀疑存在HIV-1合并感染。电泳后，提取代表异源双链的条带中的DNA，并克隆到质粒载体中；通过对一对两个混合克隆进行第二步HMA鉴定出的不同克隆进行测序，确认系统发育上不同的克隆。
在147名感染HIV-1的CSW中，有4人的自体HMA图谱与来自代表布基纳法索最常见的HIV-1分支和流行的循环重组形式（CRF）的混合参考菌株的杂交DNA的低迁移率相当。其中两人体内共存两种系统发育上不同的HIV-1群体。第一名女性呈现CRF02-AG和CRF06-cpx合并感染，第二名女性呈现CRF02-AG和一种与任何其他已知亚型无明显关系的不同病毒合并感染。在这两名女性中，通过相同的HMA程序对储存样本进行回顾性分析显示，随着血浆HIV RNA水平升高，感染了第二种病毒。
自体HMA程序结合丙烯酰胺提取异源双链，能够在一项大型队列研究中鉴定HIV-1合并感染和重叠感染。HIV-1重叠感染并非罕见现象。
