HIV抗体不确定血清蛋白免疫印迹法检测条件的正交实验优化及应用

目的优化蛋白免疫印迹法（WB）对艾滋病病毒（HIV）抗体不确定血清的检测条件,并测试其检测效能。方法采用正交实验L9（3^4）法,对HIV弱阳性对照的WB检测条件（样本量、样本孵育时间、酶联合物孵育时间）进行优化设计;对19份HIV抗体不确定血清、25份HIV阴性血清和20份HIV阳性血清,同时采用标准检测条件和优化检测条件进行检测。结果以样本量100μL、样本孵育时间2小时、酶联合物孵育时间15分钟,为HIV抗体不确定血清的最优检测条件;同标准检测条件相比,优化检测条件对HIV阴性或阳性血清的检测效能一致,但对19例HIV抗体不确定血清的检测效能更高。结论正交实验所筛选的优化检测条件可高效检测HIV抗体不确定血清。     

HIV抗体ELISA阳性标本蛋白印迹确证试验的研究

用获ＦＤＡ准许的美国ＣａｍｂｒｉｄｇｅＢｉｏｔｅｃｈ公司ＨＩＶ－１蛋白印迹试剂盒（ＷＢ）对ＨＩＶ－１／２抗体诊断试剂盒（ＥＬＩＳＡ）初筛阳性的６２份血清标本做确证试验。一次性确证ＨＩＶ抗体阳性者８名；阴性者１２名；不确定者４２名。对８名ＨＩＶ抗体不确定者做追访采样监测，其中１人１２天后ＷＢ血清ＨＩＶ抗体由不确定性转为阳性，另７人在６～１５个月经２～３次采样检测，ＷＢ区带反应无变化或略有变化。本研究证实ＷＢ不确定者，若有ＨＩＶｅｎｖ（ｇｐ１６０、ｇｐ１２０、ｇｐ４１）区带反应，ＨＩＶ血清抗体可能转阳；若仅ＨＩＶｐｏｌ（ｐ３１、ｐ５１、ｐ６６）和／或ｇａｇ（Ｐ１７、Ｐ２４、Ｐ５５）区带反应，很大可能是非特异性反应。虽ＥＬＩＳＡ阳性，而ＷＢ不确定者血清主要呈ｐ２４抗体反应（４２．９％，１８／４２）与ｐ２４、ｐ１７抗体反应（２８．６％，１２／４２）。     

菏泽市蛋白印迹试验HIV-1抗体不确定结果分析北大核心

目的分析蛋白印迹试验（WB）检测艾滋病病毒（HIV）抗体＂不确定＂结果的特征、带型特点、随访转归情况,以了解WB检测HIV抗体可能存在的问题,为HIV抗体＂不确定＂结果的随访和诊断提供参考。方法将菏泽市HIV-1抗体＂不确定＂者的检测结果、条带、社会学特征、随访及抗体转归情况录入数据库,进行整理分析。结果 21例＂不确定＂者中,自愿咨询检测（VCT）门诊检测者占47.62%（10例）,义务献血者占28.57%（6例）,住院病人占23.81%（5例）。男性占90.48%（19例）,女性占9.52%（2例）。年龄分布在17-68岁之间。21例中随访到13例,10例转为HIV抗体阳性,阳转率76.92%;3例转为阴性,占23.08%;失访7例,失访率33.33%;死亡1例。HIV抗体检测结果＂不确定＂的样本出现8种带型,分别为gp160、gp120、p24、p66、p51、p31、p17、p41,其中gp160、gp120出现频次最高。表现为12种带型组合。出现gp160/gp120带型的不确定者后期随访全部转为阳性。结论同时出现gp160和gp120两条Env带的不确定者预示有较大感染风险,对此＂不确定＂者的随访周期可缩短至1或2周。     

湖南省2008-2012年HIV抗体不确定结果分析

目的分析湖南省近5年艾滋病病毒(HIV)抗体不确定结果产生的原因和特点,为改进检测策略提供依据。方法收集2008-2012年HIV抗体不确定结果资料,结合流行病学随访、CD+4T淋巴细胞计数和病毒载量,分析不确定人群转归与蛋白免疫印迹试验(WB)带型分布的关系。结果 2008-2012年共有2995份标本进行了WB确证试验,其中不确定433例,不确定的比例为14.5%。HIV抗体不确定结果的比例逐年增长,从2008年的4.4%(21/477)上升至2012年的23.0%(182/791);其中阳性感染者比例逐年增高,尤其是晚期感染者增长迅速,分别由2009年的10.3%(3/29)和6.9%(2/29)上升到2012年的51.4%(91/177)和35.0%(62/177)。不确定结果中阴性受检者带型分布以p24或gp160为主,一般不超过两条条带;早期感染者带型分布以p24和p24+gp160为主,其中p24出现比例为100%;晚期感染者条带数目较多,显色较弱且组合更多元化,主要为p24、p31、gp120和gp160。结论随着HIV感染者增多,尤其是晚期感染比例增加,对不确定结果应予以重视,及时调整检测策略,避免漏诊。     

蛋白免疫印迹试验在HIV确证检测中的漏检及原因分析

目的探讨蛋白免疫印迹试验（WB）在艾滋病病毒（HIV）确证检测过程中的漏检问题,可能的原因及解决方法。方法 40份酶联免疫吸附试验（ELISA）和（或）胶体金法筛查阳性、WB实验确证阴性的样本,分析其来源、S/CO值,并进行抗体追踪复检。结果 40份样本中检出9份HIV阳性,28份阴性,3份样本流失。阴性者均来源于没有高危行为的普通人群,其中无偿献血人群13份,ELISA S/CO均〈3。9份阳性样本中,6例为男男性行为人群（MSM）,自愿咨询检测（VCT）3例。其中ELISA S/CO值〈6的5例,〉10的4例,6例最后考虑为窗口期感染,3例可能为WB试剂中反应膜出现问题。结论 WB检测漏检,可能是样本处于窗口期感染或试剂反应膜问题,实验室应综合分析筛查阳性而WB确证阴性的检测样本,要进行追踪检测,以确定其真实的HIV-1感染状况,防止漏检。     

220份HIV抗体检测确证为阴性或不确定标本的结果分析

目的对2007-2009年红河州220份艾滋病病毒(HIV)抗体确证为阴性或不确定的标本进行分析,以减少HIV抗体检测中假阳性结果的发生。方法按《全国艾滋病检测技术规范》(2004年)的常规HIV抗体检测方法和程序,以及HIV抗体检测的替代策略进行检测及结果判断。结果共有4 043份标本做蛋白印迹(WB)确证实验,220份标本被确定为阴性及不确定结果,占5.44(。其中阴性标本66份,占0.16%,不确定标本154份,占3.81%。结论对实验的各环节严格进行质量控制;对不确定结果的受检者应加强随访。     

3例HIV抗体WB带型结果不确定样本的检测分析北大核心

艾滋病自1981年发现以来,其检测方法和手段也在不断地改进、更新、完善。蛋白印迹法（Western blotting,WB）是确证艾滋病病毒（Human immunodeficiency virus,HIV）最常用的检测方法,确证后会出现阴性、不确定和阳性三种结果,其中不确定结果会给病人带来很大的心理压力。本文收集了3例WB带型结果首检不确定的样本,经过跟踪随访并进一步做HIV病毒载量检测,结合其血清学特点及流行病学史来探索其感染的意义,为提高HIV检测水平提供帮助。     

MIKROGEN HIV抗体条带免疫法试剂的初步评估

艾滋病病毒（Human immunodeficiency virus,HIV）抗体确认,是目前诊断HIV感染的主要实验室依据。确证试验的方法包括蛋白免疫印迹试验（Western blotting,WB）、条带免疫试验（LIATEKHIVⅢ）、放射免疫沉淀试验（Radioimmunoprecipitation assay,RIPA）及免疫荧光试验（IFA）等。国内常用的确认试验方法是WB。     

安阳市HIV抗体筛查阳性样本确认结果及条带分析

目的了解安阳市艾滋病病毒(HIV)抗体检测中,筛查试验阳性结果的准确性,以便更加合理地开展HIV抗体日常检测。方法按《全国艾滋病检测技术规范》要求进行操作,然后对比分析酶联免疫吸附试验(ELISA)筛查HIV抗体阳性者与蛋白免疫印迹试验(WB)结果的一致性。结果经两种ELISA试剂筛查HIV抗体呈阳性或一阴一阳的352份血清标本,经WB确认阳性331例,阳性率为94.03%;21例为不确定,占5.97%。WB带型≥7条带的共计328例,占99.09%;6条带的3例,占0.91%。结论艾滋病筛查实验和确认实验结果一致性高。筛查实验存在一定的假阳性,阳性样本必须进行确证实验,对确证实验不确定的样本需进行随访。     

HIV抗体ELISA阳性标本蛋白印迹确证试验的研究

用获ＦＤＡ准许的美国ＣａｍｂｒｉｄｇｅＢｉｏｔｅｃｈ公司ＨＩＶ－１蛋白印迹剂盒（ＷＢ）对ＨＩＶ－１／２抗体诊断试剂盒（ＥＬＩＳＡ）初筛阳性的６２份血清标本做确证试验。一次性确证ＨＩＶ抗体阳性者８名；阴性者１２名；不确定者４２名，对８名ＨＩＶ抗体不确定者做追访采样监测，其中１人１２天后ＷＢ血清ＨＩＶ抗体由不确定性转为阳性，另７人在６～１５个月经２～３次采样检测，ＷＢ区带反应无变化或略有变化，本研究证     

抗-HIV1+2筛查试验阳性与免疫印迹试验对比结果的分析

目的对比分析艾滋病病毒（HIV 1＋2）抗体筛查试验阳性结果与免疫印迹试验结果.方法对150份筛查复检为阳性标本的酶联免疫吸附试验（ELISA）与明胶颗粒凝集试验（PA）的结果,分别与同时检测的免疫印迹试验（WB）的结果进行比较,计算出ELISA、PA的符合率、敏感性和特异性,并依据ELISA的OD值变化探求与PA及WB结果的关系.结果ELISA、PA与WB的阳性符合率分别为89.93%和86.81%;ELISA的敏感性为100%,特异性为90.34%;PA的敏感性为100%,特异性为87.33%.凡经ELISA检测其S/CO值（平均18.63）远高于阳性对照S/CO值（平均6.677）的强阳性标本,与PA、WB的阳性符合率同为100%;经ELISA检测其S/CO值（平均0.8596）≤1或略大于1的标本,与PA、WB的阳性符合率分别为21.05%和0;经ELISA检测其S/CO值（平均2.648）＞1、小于阳性对照S/CO值的标本,与PA、WB的阳性符合率分别为66.67%和0.结论筛查试验存在一定的假阳性结果,HIV抗体结果的报告必须以WB结果为准;ELISA、PA与WB确认结果不符的情况仅出现于确认为HIV-1抗体阴性及不确定的标本中.     

622例HIV抗体筛查阳性病例采用免疫印迹试验确认结果分析

目的探讨HIV抗体ELISA筛查阳性与免疫印迹试验（WB）确证结果的关系,为HIV感染诊断提供科学依据。方法622例ELISA阳性标本与WB确证结果进行比较,采用SAS9.1进行数据描述统计和关联性分析。结果 364例确证为HIV-1抗体阳性,ELISA阳性与WB阳性符合率为58.52%。经spearman相关分析,ELISA S/Co比值与确证阳性密切相关。结论 ELISA筛查试验存在一定假阳性,高S/Co也并不代表HIV感染。     

HIV reservoirs and the possibility of a cure for HIV infection

Recent studies demonstrate that suppressive therapy can drive HIV-1 RNA levels to less than 50 copies mL(-1) in patient plasma. Yet, ultrasensitive assays show that most patients continue to harbour low-level persistent viremia. Treatment intensification studies indicate that low-level viremia could arise from several different sources. These sources include: (i) long-lived HIV-infected cells that replicate and produce virus; (ii) ongoing replication cycles in cells located in sanctuary sites where drug levels are suboptimal; and/or (iii) proliferation of latently infected cells with regeneration of a stable reservoir of slowly dividing infected cells. A well-defined latent reservoir of HIV is memory CD4+ T-cells where latency is established when an activated CD4+ T-cell becomes infected by HIV, but transitions to a terminally differentiated memory cell before it is eliminated. This review examines the dynamics and possible reservoirs of persistent HIV in patients on suppressive therapy, the mechanisms promoting viral latency and strategies to purge latent viral reservoirs. The promising research described here takes a number of steps forward to seriously address HIV remission and/or eradication.     

Broadly neutralizing antibodies suppress HIV in the persistent viral reservoir

Several highly potent and broadly neutralizing monoclonal antibodies against HIV have recently been isolated from B cells of infected individuals. However, the effects of these antibodies on the persistent viral reservoirs in HIV-infected individuals receiving antiretroviral therapy (ART) are unknown. We show that several HIV-specific monoclonal antibodies—in particular, PGT121, VRC01, and VRC03—potently inhibited entry into CD4⁺ T cells of HIV isolated from the latent viral reservoir of infected individuals whose plasma viremia was well controlled by ART. In addition, we demonstrate that HIV replication in autologous CD4⁺ T cells derived from infected individuals receiving ART was profoundly suppressed by three aforementioned and other HIV-specific monoclonal antibodies. These findings have implications for passive immunotherapy as an approach toward controlling plasma viral rebound in patients whose ART is withdrawn.The sustained suppression of HIV replication by antiretroviral therapy (ART) has dramatically improved the clinical outcome of infected individuals (1). In addition, research directed at potential pathways toward the development of an effective preventive HIV vaccine has provided insights into the nature of the immune response to HIV infection (2, 3). In this regard, recent advances in antibody-cloning technologies have led to the discovery of several highly potent and broadly neutralizing monoclonal antibodies against HIV from B cells of HIV-infected individuals (4–7). Of interest, several studies have demonstrated that certain broadly neutralizing HIV-specific monoclonal antibodies can prevent acquisition of the virus, suppress viral replication, delay and/or prevent plasma viral rebound following treatment interruption in infected animals (8–14), and block cell-to-cell transmission of laboratory-adapted HIV in vitro (15). However, it is unclear what in vivo effects these antibodies might have on HIV in humans and, in particular, what effects they may have on the virus contained in the persistently infected CD4⁺ T cells of individuals whose plasma viremia is controlled by ART. These infected CD4⁺ T cells are considered to be the major obstacle to viral eradication (16–18) as well as a potential source of plasma viral rebound following discontinuation of ART in patients whose viremia had been well controlled in therapy (1). In this regard, considerable efforts in current HIV therapeutic research have been focused on developing strategies aimed at achieving sustained virologic remission in the absence of ART (1). This focus is especially important given that viral rebound and sustained HIV replication has been observed in almost all infected individuals whose plasma viremia had been well controlled while receiving ART and whose ART was subsequently withdrawn (19). Therefore, it is important to determine which, if any, of the many recently characterized HIV-specific monoclonal antibodies can inhibit viral entry into CD4⁺ T cells of HIV isolated from the latent viral reservoir as well as replication of reservoir virus in autologous CD4⁺ T cells derived from infected individuals whose plasma viremia was well-controlled on ART. Such knowledge is critical to establishing novel opportunities for passive immunotherapy to prevent plasma viral rebound following discontinuation of antiretroviral drugs. We conducted the present study to address this issue.     

2009-2012年湖州市艾滋病确证实验室HIV抗体检测结果分析

目的分析湖州市艾滋病网络实验室艾滋病病毒（HIV）抗体检测情况和艾滋病流行特征,为指导艾滋病防治提供科学依据。方法对HIV抗体初筛阳性标本,采用酶联免疫吸附试验（ELISA）和胶体硒法进行复检,两种试剂均呈阳性或一阴一阳的用蛋白免疫印迹法（WB）进行确证,确证阳性病例进行流行病学分析。结果717份HIV抗体初筛阳性标本,复核阳性333例,确证阳性321例,其中男性230例,女性91例。各类医院检查的占45．5％（146／321）;初中及以下文化占74．1％（238／321）;异性性传播占65．1％（209／321）,男男同性性传播占15．9％（51／321）。结论应加大健康教育和对高危人群行为干预的力度,遏制艾滋病的蔓延。     

实施预防HIV母婴传播措施效果分析

目的了解实施预防艾滋病母婴传播措施对艾滋病病毒(HIV)母婴传播的影响。方法对实施干预试点地区阳性孕产妇及所生幼儿干预状况进行调查,调查幼儿出生时服药情况、喂养方式,及其母亲的分娩方式、服用抗病毒药物等情况,采用单因素χ2分析,比较干预组与未干预组幼儿HIV感染率的差异。结果实施综合干预措施组HIV母婴传播率为2.75%,没有采取任何干预措施组的母婴传播率为50.00%,两组间存在显著性差异。结论对HIV阳性孕产妇及所生婴儿及时服用抗病毒药物,并对婴儿实施纯人工喂养等措施,可有效降低HIV的母婴传播率。