HIV-1 superinfections in a cohort of commercial sex workers in Burkina Faso as assessed by an autologous heteroduplex mobility procedure
HIV-1 superinfections in a cohort of commercial sex workers in Burkina Faso as assessed by an autologous heteroduplex mobility procedure
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DOI：
10.1097/01.aids.0000131333.30548.db
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发表时间：
2004-08-20
期刊：
影响因子：
3.8
通讯作者：
Van De Perre, P
中科院分区：
文献类型：
--
作者：
Manigart, O;Courgnaud, V;Van De Perre, P
目的：描述和评估一个简单的程序，以确定HIV-1的共同或重叠感染的基础上，异源双链迁移率测定（HMA）。方法：要确定异源双链对应于不同的病毒群体在同一个人，HMA适用于单一的DNA样本，从每个主题，一个前瞻性队列的商业性工作者（CSW）在波波-迪乌拉索，布基纳法索。Env DNA扩增子变性、复性后，通过聚丙烯酰胺凝胶电泳分离杂交的DNA。通过异源双链体的缓慢迁移怀疑HIV-1合并感染，其水平与混合参考菌株相当。电泳后，提取代表异源双链体的条带中的DNA并克隆到质粒载体中;通过对两个混合克隆进行第二次HMA步骤鉴定的不同克隆进行测序，确认了系统发育不同克隆的鉴定。结果：在147例HIV-1感染的暗娼中，4例患者的自体HMA谱与来自代表最常见HIV的混合参考菌株的杂交DNA的低迁移率相当，1进化枝和循环重组形式（CRF）流行于布基纳法索。在其中两个国家，两种遗传学上不同的HIV-1人群共存。第一名女性出现CRF 02-AG和CRF 06-cpx合并感染，第二名女性出现CRF 02-AG和与任何其他已知亚型无显著相关的趋异病毒合并感染。在这两名妇女，回顾性分析存储样品相同的HMA程序显示收购的第二个病毒伴随着增加血浆HIV RNA.Conclusions：AutoblastHMA程序，然后由丙烯酰胺提取异源双链体允许识别HIV-1的共同和超级感染在一个大型队列研究。HIV-1重叠感染并不是一种罕见的现象。(C)2004年利平科特威廉姆斯威尔金斯。
Objective: To describe and evaluate a simple procedure to identify HIV-1 co- or super-infections based on a heteroduplex mobility assay (HMA).Methods: 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.Results: 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.Conclusions: 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. (C) 2004 Lippincott Williams Wilkins.