景德镇市490例HIV-1抗体阳性者免疫印迹带型分析

目的对景德镇市490例HIV-1抗体阳性者的免疫印迹带型进行分析。方法采用蛋白免疫印迹试验对标本进行确证检测,分析不同性别、不同年龄组的带型差异。结果 gp160、gp120、p66、p55、p51、gp41、p39、p31、p24、p17全部阳性的共142例,条带gp160、gp120出现率最高为100%,p24为99.2%,gp41为95.7%,p55出现率最低为43.9%;条带gp41和p17在男女性别中的出现率差异有统计学意义(P0.05),条带p31在不同年龄组间的出现率差异有统计学意义(P0.05)。结论gp160、gp120、gp41、p24是判定HIV-1抗体阳性的重要条带,本市发现的HIV感染者多数在AIDS期前。     

54例HIV抗体确认阳性无偿献血者蛋白印迹实验带型分析

目的对本地区HIV抗体确认阳性献血者的带型特征进行分析,以对献血招募、职业防护及血液检测提供理论指导。方法经本中心实验室初筛检测为HIV抗体反应性标本送市疾控中心进行蛋白免疫印迹法(western blotting,WB)确认检测,收集经确认实验为HIV抗体阳性的标本信息,对各条带出现率及条带组合情况分别进行汇总分析。结果共收集HIV抗体确认阳性献血者54例,带型以全带型比例最高,为38.9%;各条带检出率:gp160/120、p24均为100%,其次为p66、gp41分别为92.6%、90.7%,p31、p17、p51依次为83.3%、87.0%、87.0%,p39和p55阳性率最低,分别为59.2%、42.6%。结论带型分析显示HIV抗体确认阳性献血者以早期感染为主,应采取措施避免高危人群献血。     

抗HIV抗体免疫印迹法检测中阳性及不确定结果的带型分析

目的分析抗HIV抗体免疫印迹(western blot,WB)试验检测中出现的阳性和不确定结果的带型特征,探讨WB带型与HIV感染的关系。方法收集近5年检测的189例抗HIV抗体阳性样本及14例抗HIV抗体不确定样本,分析WB确证试验的带型特征。结果在189例抗HIV抗体阳性的样本中,出现WB全带型(gp160,gp120,p66,p55,p51,gp41,p31,p24,p17均阳性)共80例(占42.33%)。其中gp160、gp120条带阳性率均为100%,p55条带阳性率最低,为40.74%。14例不确定样本中共有5种带型,gp160、p24(±)带型9例(占64.29%),p24带型2例(占14.29%),gp160,gp160、gp120(±)和p24、p17带型各1例。9例gp160、p24(±)带型中,2例复检转为阳性。结论抗HIV抗体阳性样本WB带型以全带型为主。抗HIV抗体不确定样本以gp160、p24(±)带型为主。出现此带型时应加强随访以免漏检。     

不同临床分期HIV-1感染者/AIDS患者血样的蛋白印迹试验带型分析

目的分析各临床分期的人类免疫缺陷病毒1型(HIV-1)感染者/获得性免疫缺陷综合征(AIDS)患者血清中病毒蛋白抗体的变化,探讨不同临床分期的HIV-1感染者/AIDS患者蛋白印迹试验(WB)的带型特点。方法对汕头市2009至2012年确证的部分HIV-1抗体阳性病例进行CD4+T细胞计数,根据计数结果对病例进行分组:原发感染期组(CD4+≥500/mm3);感染中期组(CD4+≥200/mm3且500/mm3);AIDS期组(CD4+200/mm3);对各组的WB结果进行病毒蛋白(p17、p24、p31、p39、gp41、p51、p55、p66、gp120、gp160)抗体的阳性率统计以及带型分析。结果病毒蛋白p24、p31、gp41、p51、p66、gp120和gp160的抗体阳性率较高(总阳性率分别为98.2%、92.0%、90.8%、89.5%、93.3%、98.7%和97.5%),且在3个分期组间未见明显变化(P0.05);病毒蛋白p17、p39和p55呈现较低的抗体阳性率(总阳性率分别为68.9%、49.1%和42.5%);蛋白p17的阳性率从原发感染期的91.5%降为感染中期的62.8%(P0.01)及AIDS期59.3%(P0.01);WB结果中的全带型、缺失p39+p55和缺失p17+p39+p55的带型是3个疾病分期组患者的常见带型;在3个分期组中,原发感染期中全带型出现率最高(60.8%),而缺失p17+p39+p55的出现率最低(6.5%)。结论病毒蛋白p17抗体的阴转可以作为疾病从原发感染期进入感染中期/AIDS期的一个潜在判别指标。     

利用logistic回归模型对艾滋病病毒抗体不确定者转归分析

目的利用logistic回归模型,分析艾滋病病毒（HIV）抗体不确定者的初筛、免疫印迹试验（WB）条带结果和其随访阳转的关系,提高对转归的预判断。方法收集2012 — 2016年HIV抗体不确定者酶标、硒标结果、WB条带结果,用受试者工作特征（ROC）曲线确定酶联伯乐试剂阈值,用logistic回归筛选WB条带的危险因素并建立预测模型。结果79份不确定标本中,首次2种初筛结果都阳性与最终随访确证阳性符合率高达96.29%。 ROC曲线确定伯乐试剂的特定阈值为8,S/CO＞8提示随访转归阳性概率大。最常见的带型分别为p24、gp160＋p24和gp160＋gp120＋p24;其中p24带型随访阴性的比例最高,占77.27%,gp160＋p24有或无其他条带随访阳性概率较大。 运用logistic回归模型筛选出gp160、p24、p17三个有意义的因素。结论初筛酶标、硒标检测结果双阳性,S/CO＞8,gp160＋p24有或无其他条带,随访结果为阳性概率较大。 联合初筛结果、带型情况可较为客观、准确地预测HIV不确定患者转归结果。     

核酸检测技术在献血人员血液筛查中的应用

目的：采取核酸检测（NAT ）技术进行血液筛查,减少由献血者感染“窗口期”及隐匿感染献血引起的疾病传播。方法采用酶联免疫吸附法（ELISA）对54358份献血者标本进行乙型肝炎表面抗原（HBsAg）、人类免疫缺陷病毒抗体（抗-HIV）、丙型肝炎病毒抗体（HCV-Ab）检测,同时用转录介导扩增技术（TMA）进行 HBV-DNA 、HCV-RNA 检测,人类免疫缺陷病毒1型（HIV-1）RNA 核酸检测。结果 NAT 检测阳性 ELISA 检测阴性标本共51份。51份标本有鉴别试验结果的33份,其中32份 HBV-DNA 阳性,1份 HIV-1 RNA 阳性,HIV-1 RNA 阳性的献血者经南京市疾病预防控制中心免疫印迹法确认为 HIV-1疑似阳性（gp160和 p24±）。半年后再次回访该献血者,抽取血样送南京市疾控中心经免疫印迹法确认为 HIV-1阳性（gp160、gp120、p66、p51、gp41、p31、p24、p17＋）。结论 NAT 技术可以减少由献血者感染“窗口期”及隐匿感染献血引起的疾病传播,从而减少输血风险。     

抗-HIV抗体筛查与免疫印迹试验两种检测方法的对比研究

目的探讨了酶联免疫吸附试验(ELISA)法和免疫印迹试验(WB)两种方法检测HIV抗体的准确性。方法用ELISA方法检测了7 291例样本,对41例复测阳性的标本用WB法检测,分析两种方法检测结果的关系。结果 41例阳性样本用WB法检测,共检出阳性样本12例(S/CO平均值为22.8);阴性结果13例(S/CO平均值为8.7),不确定结果16例(S/CO平均值为10.7)。阳性标本中p55条带出现的频率为47.1%,p51为58.3%,其他均达到75.0%以上。结论筛查试验存在假阳性结果,HIV抗体先筛查,后确证有利于提高HIV抗体检测结果的准确性。     

液相芯片法验证蛋白质免疫印迹试验检测HIV-1结果的敏感性和特异性

目的初步分析采用液相芯片法研制人类免疫缺陷病毒(HIV)-1检测试剂的可行性。方法采用液相芯片技术用蛋白质免疫印迹试验(Western Blot)检测HIV-1阳性和阴性的标本;用HIV-1的gp120、gp41、p24抗原分别包被不同编号的免疫磁珠,与生物素化二抗、亲和素化荧光染料PE组成液相芯片检测系统,对标本进行检测分析。结果 55份标本经Western Blot法确认阳性样本检测符合率为100%;76份经酶联免疫吸附试验(ELISA)初检和复检阴性的标本检测HIV-1gp120、HIV-1gp41、HIV-1p24抗原均为阴性;86份经ELISA法检测初检和复检为阳性而Western Blot法确认为阴性的样本,其HIV-1gp120、HIV-1gp41、HIV-1p24单片段检测均为阴性。液相芯片法检测HIV-1抗体的敏感度和特异度均为100%。结论采用液相芯片技术组成的HIV-1抗体检测系统,其敏感度和特异度均优于ELISA法检测。     

HIV抗体检测结果不确定病例的核酸定量检测分析

目的 初步探讨HIV抗体不确定人群的带型特征以及核酸定量检测结果与阳转结果的关系。方法 收集2015—2020年南昌市第九医院初次检测结果为HIV抗体不确定的142份标本。用酶联免疫吸附实验（ELISA）、蛋白印迹试验（WB）实验以及核酸定量检测方法对上述HIV抗体不确定标本进行检测,以随访结果为判断标准,比较分析不同检测方法结果和抗体转归的相关性。结果 142例随访者中,有85例出现血清学阳转,占比59.86%（85/142）。阳转者免疫带型gp160出现比率为97.64%（83/85）,p24出现比率为49.41%（42/85）;未阳转样本gp160出现比率为2.36%（2/85）,p24出现比率为50.59%（43/85）。初次检测抗体不确定检测样本中gp160条带出现率在阳转者和未阳转者之间的比较有统计学差异（χ2=49.41,P<0.01）;而二者p24条带出现率比较,差异没有统计学意义（χ2=2.05,P>0.05）。核酸检测试验结果显示83例抗体不确定样本呈核酸阳性,核酸阳性与抗体阳转的符合率为97.64%（83/85）。结论 HIV抗体不确定样本中出现gp160带型者随访转归为阳性的可能较大;核酸检测试验可为抗体不确定样本提供进一步鉴别的依据。     

蛋白印迹法检测津巴布韦HIV患者

津巴布韦血液中心用蛋白印迹法(WestemBlot,WB)检测HIV阳性或阴性者288例,所有患者均已用Abbott重组酶免疫法(REIA)检出有HILV-Ⅲ抗体。 45份REIA法阴性标本,经WB法检测,44份仍为阴性(无带);1份未能确定(弱P24,P51,P66和P160带)。62份REIA法HIV低滴度标本(报告为可疑的/未能确定的HIV抗体)经WB法试验,28份为阳性(全带),30份为阴性(无带);4份不能确定(1份为仅有gp41带的低滴度HIV抗体,3份为P24带的     

57例抗HIV初筛阳性确认实验带型分析

目的 调查HIV感染初筛实验的准确率及确认抗HIV初筛试验阳性血清的感染和感染型别。方法 调查我国艾滋病高发农村，对部分初筛阳性和可疑的血清进行确认实验。结果 57例初筛为抗HIV阳性和可疑血清进行确认实验，56份为HIV—l阳性，其中1份样本出现HIV—2型反应条带；1份阴性。确认阳性标本的反应条带中，抗外膜蛋白(env)抗体阳性率最高，gpl60为100％，gpl20为94．6％，gp41为91．1％；多聚酶抗原(pol)p66为82．1％；核心抗原(gag)P24为53．6％。提示无症状携带组确认反应条带的pol和gag的阳性率显著高于艾滋病组(P＜0．01)。儿童组gag阳性率显著低于成人组(P＜0．05)。结论 HIV感染初筛实验的准确率较高，但确定感染需靠确认实验。外膜蛋白、p66和p24抗原是HIV感染的重要抗原，对确认HIV的感染具有指导意义。     

Serologic markers for staging human immunodeficiency virus infections

In the present study, 80 serum specimens were tested for human immunodeficiency virus (HIV) antibody by enzyme immunoassay (EIA) and indirect fluorescent antibody tests, HIV (p24) antigen, and Western Blot analyses. A total of 40 specimens were HIV antigen-positive, 35 were antigen-negative and five were indeterminant. Among the 40 antigen-positive sera, 38 had positive antibodies by EIA with confirmation by Western Blot. Two cases were antigen-positive and were thought to be early stages where antibodies had not yet developed. Among the 38 sera, 30 (79%) had decreased or had no reactions by indirect fluorescent antibody tests. Among the 35 antigen-negative cases, all 35 had positive antibodies by EIA and all 35 had bands at gp41 by Western Blot. Among 84 HIV-infected patients, 30 had >400 CD4+ cells per cubic millimeter, 21 patients had 200–400 CD4+ cells and 33 had <200 CD4+ cells. A total of 28 (93%) of the HIV antigen-negative cases with full banding patterns by Western Blot had >400 CD4+ cells. In contrast 18 (55%) of the patients with antigen positivity and incomplete banding on Western Blot had <200 CD4+ cells.     

深圳东部地区无偿献血人群中HIV感染者特征及蛋白质免疫印迹带型分析

目的掌握深圳东部地区无偿献血人群中HIV感染者人口学特征及蛋白质免疫印迹(WB)带型,为献血者招募、员工职业防护和血液检测提供理论依据。方法收集2010—2019年深圳东部地区无偿献血人群经WB法确认的62例HIV感染者资料,采用χ²检验比较各年度感染率;对HIV感染者的性别、年龄、户籍、职业、文化程度、婚姻状况构成比进行描述性统计分析;对WB条带阳性率和带型分别进行汇总分析。结果深圳东部地区2010—2019年无偿献血者HIV的总感染率为3.92。各年度献血者的HIV感染率比较,差异有统计学意义(χ²=29.44,P<0.05)。2019年A采血点的感染率为12.45,与其他采血点总感染率(1.81)比较,差异有统计学意义(χ²=5.88,P<0.05)。62例感染者中男性占93.55%;18~<36岁的感染者占74.20%;未婚感染者占61.29%;非深圳户籍人群占93.55%;商业服务人员和工人分别占43.55%、32.26%;大专以下学历占88.71%。WB带型以4种带型为主;条带gp160、gp120、p66、p24阳性率均为100.00%,p17、p55阳性率较低,分别为67.74%、51.61%。结论深圳东部地区无偿献血者HIV感染者以早、中期感染的男性、18~<36岁、未婚、非深圳户籍、商业服务人员或工人、大专以下学历人群为主,尤其要关注A采血点的献血人群。     

Urinary HIV-1 antibody patterns by western blot assay.

Diagnosis of human immunodeficiency virus infection (HIV-1) is normally carried out by serum testing for HIV-1 antibody. Recently, antibody testing in other body fluids such as saliva and urine have been attempted. In this study, we examined HIV-1 antibody patterns in urine by Western blot assay as compared to that found in serum. Out of 44 sero-positive samples by Western blot assay we found 43 to be HIV-1 antibody positive in the urine, whereas all 40 sero-negative samples were negative in urine. Thus the sensitivity of urine testing was 97.7% with 100% specificity when compared to serum testing by the Western blot assay. In the analysis of the antibody pattern in urine, we found 6.8% of p17, 68% of p24, and 47.7% of p39 in the core proteins; 72.7% of p31, 61.4% of p51, and 68.2% of p66 in the polymerase; and 63.6% of gp41, 75% of gp120, and 97.7% of gp160 in the envelope proteins. The data obtained supports the selection of the HIV-1 antigen subtype-E to develop a home test kit using urine. Urine testing for HIV-1 antibody is convenient, non-invasive, safe, and easily performed at home. However, if the urine is positive, the confirmation test on serum is needed.     

太原市无偿献血者抗-HIV结果确认和带型分析

目的分析研究太原市无偿献血者人类免疫缺陷病毒(HIV)抗体(抗-HIV)阳性标本的确认结果和带型。方法采用蛋白印迹(WB)对185例初筛阳性献血者标本进行确认试验,采用SPSS13.0分析试验结果和带型。结果确认阴性128例,不确定15例、阳性42例。确认阳性标本中,gp120、gp160、p24带型阳性率为100.00%,p31、p51和p66阳性率均大于90.00%。不确定标本中,p24阳性率最高,有12例,占80.00%;其次为gp160,有3例,占20.00%。随着S/CO值增大,带型出现率逐渐增高,最高为S/CO值6.00组(χ~2=35.16,P=0.009)。结论 HIV筛查试验存在假阳性。确认阳性标本多为病毒感染期。初筛阳性标本必须进行追踪确认试验,确认其是否感染。     

False-positive human immunodeficiency virus type 1 western blot tests in noninfected blood donors

Background: The manufacturers' criteria for a positive human immunodeficiency virus type 1 (HIV-1) Western blot (WB) test were recently revised to require reactivity to only two of the following bands: p24, gp41, and gp120/160. In a recent report, low-risk blood donors were identified in whom nonspecific reactivity to multiple env antigens in WB testing resulted in apparently false-positive WBs by these criteria. The present study was conducted to verify the existence of false-positive WBs among noninfected donors and to assess the extent of this problem. Study Design and Methods: Four donors classified as WB- positive on the basis of env-only (3 cases) or p24/env-only (1 case) patterns were investigated. Index and/or follow-up specimens were tested by polymerase chain reaction (PCR), by overlapping recombinant env antigens and synthetic peptides in enzyme immunoassays, and by deglycosylated and denatured antigen WBs. WB records from American Red Cross blood centers were reviewed to determine the frequency of env- only and p24/env-only patterns, relative to all positive WBs, from 1988 through 1993. Results: The four index-case donors denied risk and had stable WB reactivity during follow-up. HIV PCR was negative in all. Env reactivity was restricted to nonglycosylated gp41 epitopes; no gp120- specific reactivity was detected. For three of the four donors, env reactivity was mapped to a 20-amino acid N-terminal epitope of gp41. The rate of detecting WBs with these false-positive patterns increased from 0.6 percent of all positive WBs from 1988 to 1990 (4/776) to 8 percent in 1991 and 1992 (52/683), and then it declined to 6 percent in 1992 and 1993 (47/783). Env-only patterns predominated in 1991 and 1992, whereas p24/env-only patterns were more frequent following implementation of combined anti-HIV-1/HIV type 2 enzyme immunoassays in 1992. Conclusion: Low-risk blood donors can have false-positive results on WB tests. Increased detection of env-only and p24/env-only WBs appears related to the enhanced sensitivity of newer enzyme immunoassays to gp41 and p24 antibodies. Donors with these patterns should undergo follow-up testing to document the presence or absence of HIV infection.     

Conversion of an immunogenic human immunodeficiency virus (HIV) envelope synthetic peptide to a tolerogen in chimpanzees by the fusogenic domain of HIV gp41 envelope protein

The fusogenic (F) domain of human immunodeficiency virus (HIV) gp41 envelope (env) protein has sequence similarities to many virus and mediates the fusion of HIV-infected cells. During a survey of the immunogenicity of HIV env peptides in chimpanzees, we have observed that HIV peptide immunogenicity was dramatically altered by the NH2- terminal synthesis of the gp41 F domain to an otherwise immunogenic peptide. We compared two hybrid peptide types comprised of T helper (Th) and B cell epitopes of HIV gp120 env protein for their immunogenicity in chimpanzees. The Th-B epitope hybrid peptides contained the HIV gp120 Th cell determinant, T1 (amino acids [aa] 428- 440)-synthesized NH2 terminal to gp120 V3 loop peptides, which contain B cell epitopes that induce anti-HIV-neutralizing antibodies (SP10IIIB [aa 303-321] and SP10IIIB [A] [aa 303-327]). The F-Th-B peptide contained the HIV gp41 F domain of HIVIIIB gp41 (aa 519-530)- synthesized NH2 terminal to the Th-B peptide. Whereas Th-B peptides were potent immunogens for chimpanzee antibody and T cell-proliferative responses, the F-Th-B peptide induced lower anti-HIV gp120 T and B cell responses. Moreover, immunization of chimpanzees with F-Th-B peptide but not Th-B peptides induced a significant decrease in peripheral blood T lymphocytes (mean decrease during immunization, 52%; p < 0.02). Chimpanzees previously immunized with F-Th-B peptide did not respond well to immunization with Th-B peptide with T or B cell responses to HIV peptides, demonstrating that the F-Th-B peptide induced immune hyporesponsiveness to Th and B HIV gp120 env determinants. These observations raise the hypothesis that the HIV gp41 env F domain may be a biologically active immunoregulatory peptide in vivo, and by an as yet uncharacterized mechanism, promotes primate immune system hyporesponsiveness to otherwise immunogenic peptides.     

Reliable confirmation and quantitation of human immunodeficiency virus type 1 antibody using a recombinant-antigen immunoblot assay

The recombinant DNA-derived, human immunodeficiency virus (HIV) antigen-based immunoblot assay (RIBA-HIV216) is a new supplemental (confirmatory) test developed to detect antibodies to HIV-1. The assay employs four recombinant viral antigens, corresponding to HIV-1 p24, p31, p41 and gp120 proteins, in an immunoblot format. With this assay, HIV-1 antigen reactivity was detected in all 683 infected patient serum or plasma specimens evaluated; 665 (97.6%) of these sera met the criteria for a positive interpretation, and 18 (2.6%) were classified as indeterminate. All 683 samples reacted with the recombinant gp41-equivalent protein. The first sequential enzyme immunoassay (EIA)-reactive samples collected from 33 seroconverting homosexual men reacted on RIBA-HIV216. Eleven (1.1%) of 999 EIA-negative blood donor sera reacted weakly with a single recombinant antigen (p24 or p31), whereas 13 to 48 percent had indeterminate reactions on viral lysate Western blots. One (1.5%) of 66 EIA-positive, Western blot-negative blood donor samples and 19 (29%) of 66 EIA-positive, Western blot-indeterminate blood donor samples scored indeterminate on RIBA-HIV216. Nonspecific reactivity was seen with only 1 (0.8%) of 114 patient sera containing possible interfering antibodies, whereas 33 percent of these samples had indeterminate reactions on Western blot and 35 percent had equivocal reactions on immunofluorescence assay (IFA). We conclude that the RIBA-HIV216 is approximately as sensitive as and significantly more specific than virus-derived Western blot and IFA. The RIBA-HIV216 also allows for semiquantitation of specific antibodies that may be of value in clinical staging and therapeutic monitoring.     

Incidence of anti HIV antibodies and viral antigen in standard and control sera

Most material used for control and calibration in a clinical laboratory is based on pool sera of human origin, guaranteed to be HBsAg-free. Since little information is available on the potential infectivity of HIV, the causative agent of acquired immunodeficiency syndrome (AIDS), 54 control and calibration sera, in routine use, were investigated for the incidence of antibodies to HIV by means of Elisa. Sixteen test specimens ( = 30%) gave positive or borderline Elisa results and were further analysed by immunoblotting, resulting in 15 samples all recognizing gp 160 and partially the p 24, p 31, p 55, p 64 and gp 120 band. Only one sample with borderline Elisa result was negative by this assay. Furthermore, all sera were examined for the presence of viral antigen by a solid phase Elisa. All samples under investigation gave negative antigen Elisa results. Bearing in mind that the sensitivity of this assay is limited to 10 micrograms/l of viral antigen, no conclusion on infectivity should be drawn. The high incidence of HIV-antibodies in the sera investigated demands that this material should be handled with special care by laboratory personnel.