血清HIV抗体阴性静脉药瘾者肝组织HIV DNA测定.docVIP

血清HIV抗体阴性静脉药瘾者肝组织HIV DNA测定 血清HIV抗体阴性静脉药瘾者肝组织HIV DNA测定 2010-01-11 吴南屏　刘克洲　陈智　李继承　R.H.Dennin 　　【摘要】　目的　了解血清抗-HIV阴性的静脉药瘾者肝组织是否存在前病毒HIV DNA。方法　 选择38例抗-HIV阴性的静脉药瘾者肝组织，10例非静脉药瘾者肝组织，用巢式聚合酶链反应，分别在HIV gag、pol、env三大结构区，用六对引物进行HIV DNA扩增。结果　38例中有3例同时在三个区域测到HIV DNA，12例分别在二个区域、17例在单一区域检测到HIV DNA，6例在HIV三个区域均阴性。对照组10例无一例检到HIV DNA。结论　抗-HIV阴性的静脉药瘾高危人群，存在潜在HIV感染。提示潜在HIV感染的检测及加强“窗口期”HIV的监测十分重要。 　　【关键词】　静脉药瘾　　抗-HIV　　人类免疫缺陷病毒DNA Detection of HIV DNA by nPCR in liver tissue of anti-HIV seronegative intravenous drug users　Wu Nanping,Liu Kezhou,Chen Zhi,et al.　Institute of Infectious Diseases,Zhejiang Medical University,Hangzhou 310003 　　【Abstract】　Objective　The study was designed to investigate whether there existed provirus HIV DNA in the liver tissues of intravenous drug users (IVDU) with seronegative results of anti-HIV test.Methods　We detected 38 liver tissues of seronegative IVDUS and 10 liver tissues of non-IVDUS.Using different primers in HIV gag,env as well as pol regions respectively,HIV DNA was detected by nPCR.Results　In all 38 cases,3 simultaneously reacted to all three HIV regions (HIV,gag,env,pol),12 reacted to two HIV regions,17 reacted to single HIV region and 6 cases did not react to any HIV region.No sample of control group was found reacting to any HIV region.ConclusionIt showed that there might exist HIV infection is some of HIV seronegative IVDUS at high risk of HIV infection.So it is very important to detect potential infection and prevent “windows” HIV infection. 　　【Key words】　Intravenous drug users　　Anti-HIV　　HIV DNA 　　从HIV感染到机体产生抗-HIV，其“窗口期”时间长短直接与HIV感染的量、毒力、感染方式及人体遗传、免疫状况有关。国外报道已在血清HIV抗体阴性的静脉药瘾者中找到特异的HIV DNA，其阳性率高达23%～80%［1～6］，但作者尚未见组织中找到HIV DNA的报道。我们对38例抗-HIV阴性静脉药瘾者肝组织，用六对引物分别在HIV LTR-gag、pol、env区进行HIV核酸扩增，以了解高危人群HIV抗体阴性者HIV感染情况及探讨可能的HIV免疫逃逸机制。 材料与方法 　　一、标本来源 　　38例标本均来自德国，为血清抗-HIV阴性的静脉用药过量死亡者的肝组织。无菌条件下取肝组织，迅速深低温-70℃保存备用。死亡前作为HIV高危人群，ELISA(德国Mercke公司)测定为抗-HIV阴性。10例对照组标本系抗-HIV阴性的因交通事故经外科手术切除的部分肝组织标本。 　　二、肝组织DNA抽提 　　取400mg新鲜冻存肝组织，低温下磨碎(匀浆器事先用三蒸水处理180℃30