间接免疫荧光试验在HIV抗体检测中的应用

目的：评价间接免疫荧光试验（IFA）作为一种确认实验在HIV抗体检测中的应用。方法：用T嗜性的HIV毒株感染MT4细胞制备抗原片,并与不同稀释度的免疫血清相结合,再滴加荧光素标记的羊抗人IgG,在荧光显微镜下检测特异性免疫荧光,结果：用IFA检测43分血清样本（其中包括17份WB阳性血清,14份阴性血清和12份ELISA阳性或WB出现可疑条带但按标准不能判为阳性的可疑血清）,除HIV－2血清呈阴性反应外,其余检测结果与WB相符,IFA对HIV－1抗体检测的敏感性和特异性均达到100％,HIV－1阳性血清抗体最高滴度达到1：10240。结论：IFA可用于HIV－1抗体检测的确认,它可作为Western Blot的一种补充或替代实验。     

广州市发现首例HIV-1/HIV-2混合感染

目的探讨首例HIV-1/HIV-2混合感染发现的经过以及对本地艾滋病预防控制的意义。方法对广州市某医院一名门诊患者血样2份(包括初筛试验前后各1份)进行HIV抗体检测。初筛试验采用酶联免疫吸附试验(ELISA)和明胶颗粒凝集试验(PA),确认试验采用蛋白印迹法(WB)。结果患者为男性,非洲马里人,血清经初筛试验为HIV阳性。初筛阳性的血清,以HIV-1/HIV-2 HIVBLOT2.2试剂做蛋白印迹试验(WB),确认为HIV-1阳性,同时HIV-2的特异条带gp36阳性;样本再用HIV-2型的BLOT1.2膜条确认为HIV-2阳性。结论被检者为HIV-1/HIV-2混合感染,这是广州市、广东省首次发现HIV-1/HIV-2混合感染,提示HIV-2可能会以对外交流等方式传入广东。     

HTLV-I and HTLV-II infections among HIV-1 seropositive patients in Sao Paulo,Brazil

To estimate the presence of, and the risk factors for HTLV-I and HTLV-II infections among HIV-1 infected subjects in Sao Paulo, Brazil, a serosurvey was performed in 471 HIV-1 infected patients, including 216 intravenous drug addicts (IVDA), 229 homosexual/bisexual men, and 26 with other risk factors. Serum samples were screened for HTLV seroreactivity by ELISA; reactive samples were analyzed by Western Blot (WB), using whole HTLV-I lysate as antigen. To confirm and discriminate HTLV-I and HTLV-II infections, sera presenting any bands on WB were further analyzed by a WB containing recombinant HTLV-I and HTLV-II proteins (WB 2.3), and by enzyme immunoassays using synthetic peptides specific for envelope proteins (Synth-EIA). In 22 cases, cell samples were available for polymerase chain reaction (PCR) studies. On WB, 114 sera were reactive and, of these, 37 and 25 were concordantly positive on both WB 2.3 and Synth-EIA procedures for HTLV-I and HTLV-II specific antibodies, respectively; 37 specimens were negative on both assays, and 15 gave discordant or indeterminate results. PCR findings confirmed concordant results obtained in the discriminatory serological assays. The prevalence rates of HTLV-I and HTLV-II infections were 15.3% and 11.1% in IVDA, and 0.9% and 0.4% in homosexual/bisexual men, respectively. No case of HTLV-I/HTLV-II co-infection was found.     

我国人类免疫缺陷病毒2型感染的初步回顾性血清学调查

[目的]初步了解我国人类免疫缺陷病毒2型（HIV-2）的感染状况。[方法]用多种HIV抗体确认方法检测来自全国各地的HIV-2可疑血浆/血清样品,用Genelabs HIV Blot2.2WB检测5份已知HIV-2血浆样品。[结果]用Genelabs HIV Blot2.2WB检测54份HIV-2可疑样品,全部呈HIV-1抗体阳性反应且有HIV-2指示带;用HIV-2WB试剂检测,44.4%（24/54）呈HIV-2抗体阳性反应、55.6%（30/54）呈不确定反应;用2种线性免疫试验检测,分别只有3.7%（2/54）和1.9%（1/54）判定为HIV-1/2混合感染,绝大部分仅为HIV-1抗体阳性。对于5份已知HIV-2样品,使用Genelabs HIV Blot2.2WB检测时与HIV-1抗原发生不同程度的交叉反应,但带型明显不同于HIV-1样品。[结论]结果提示我国的HIV-2感染很少见,现有的HIV-2WB不宜用于检测HIV-1抗体呈阳性反应的样品。     

One year surveillance of HIV-1 infection in Johannesburg, South Africa

A sero-epidemiological surveillance study to monitor the prevalence of HIV-1 infection in Johannesburg, South Africa, was commenced in February 1988. The population selected for study were attenders at clinics for sexually transmitted diseases (STD) and at family planning (FP) clinics. In the 12 months of the study 6631 sera were tested. Of the STD attenders, 15 of 1224 black females (1.2%) and 21 of 2482 black males (0.8%) were positive. Of the 449 white males tested 49 were homosexual, amongst whom 10 (20.4%) were positive; in the heterosexual white male group 4 of 400 (1.0%) were positive. Of the FP clinic attenders, 4 of 1459 black females (0.3%) were positive. 68 of the 6631 sera tested were indeterminate for infection. No attenders were positive for HIV-2 infection. These data confirmed the entry of HIV infection into the black population in South Africa.     

Assessment of serological assays for the diagnosis of HIV-1 infections.

Many serologic techniques are available for the detection of HIV-1 infection, but their diagnostic performances have been reported to vary depending on the testing situations. We evaluated eight different serologic assays for detecting HIV-1 antibody in sera from suspected high risk groups in North and East Africa. These included: one ELISA bead assay, three indirect ELISAs, two competitive ELISA assays, one gelatin agglutination assay and a recombinant dot-blot assay. A panel of 38 HIV-1 antibody confirmed positive human sera and a panel of 60 HIV negative sera were tested. All sera yielding discordant results were retested for verification of reactivity. Among the different assays evaluated for detecting HIV-1 antibody, sensitivities ranged from 0.919 to 0.974 and test efficiencies ranged from 94.2-98.9%. All assays failed to detect at least one western blot confirmed positive serum; three tests (Organon, DuPont recombinant and Serodia) produced only one false-negative result. Five of the eight assays did not produce any false-positive results. The Organon viral lysate and the DuPont recombinant ELISAs exhibited the best overall performance (test efficiency = 98.9%).     

医院就诊者HIV抗体结果分析

目的:分析医院就诊者人艾滋病病毒(HIV)抗体检测情况,为医院预防艾滋病提供依据。方法:2004年至2011年,分别用中山、梅里埃、丽珠三种ELISA试剂和雅培硒标试剂筛查HIV抗体,筛查阳性样本送广州市疾病预防控制中心用WB法进行确证。结果:333 472例各种疾病患者中筛查出阳性样本564例。确证阳性332例,流行率0.100%,2006年-2011年合计显著高于2004年-2005年合计(χ2=12.65,P=0.000)。以18岁～50岁的青壮年男性居多。HIV感染者广泛分布于医院的40个临床科室,门诊180例(54.22%);住院152例(45.78%),其中内科131例,外科仅21例。确证阴性192例。18个月以下婴幼儿待确证4例。确证不确定36例,随访后2例证实感染HIV、5例转阴性、1例仍为不确定,其余失访。结论:医院就诊者HIV感染率高于中国全人群感染率,感染者广泛就诊于各个科室,建议将HIV抗体检测常规化。医院在发现HIV感染者方面发挥了重要作用。     

一种血液艾滋病病毒抗体免疫印迹试剂的现场评估

目的对Cambridge血液艾滋病病毒（HIV）-1抗体免疫印迹（WB）试剂进行现场评估并考核其在不同的HIV感染状态的人群中确认试验检测的敏感性和特异性.方法选取不同的现场,分别采集不同的HIV感染状态人群的血液样品,共计645份.经酶联免疫吸附试验（ELISA）检测后,分别使用Cambridge血液HIV-1抗体WB试剂盒和HIV blot 2.2 HIV-1/2型WB确认试剂盒进行平行对比试验.结果在已知既往HIV感染者中,Cambridge血液WB与Genelabs WB检测结果均为阳性,在此人群中上述2种确认试剂的敏感性均为100%.在398例HIV抗体ELISA检测为阴性的人群中,Cambridge血液WB试验23例为不确定;Genelabs血液WB试验86例为不确定,在此类人群中上述2种确认试剂的特异性分别为94.22%（375/398）和78.39%（312/398）.结论Cambridge血液HIV-1抗体WB试剂盒与Genelabs诊断公司的HIV blot 2.2 HIV-1/2型WB试剂盒的试验结果对比,在特异性方面前者优于后者.     

HTLV-1, HIV-1, hepatitis B and hepatitis delta in the Pacific and South-East Asia: a serological survey

Blood samples from 13 locations in the Pacific and South-East Asia were tested for evidence of infection with human T-cell lymphotropic virus type-1 (HTLV-1), human immunodeficiency virus (HIV-1), hepatitis B virus (HBV) and hepatitis delta virus (HDV). No samples were positive for antibody to HIV-1. Antibodies to HTLV-1 were found in samples from five locations, the maximum prevalence being 19%, in Vanuatu. Serological markers of HBV infection were found in all locations, the maximal prevalence being 88%, in Majuro, Micronesia. Antibodies to HDV in HBsAg positive sera were found in six locations with a maximum prevalence of 81% in Kiribati.     

2013年HIV抗体免疫印迹试验确证不确定结果分析

摘要:目的　分析人类免疫缺陷病毒（HIV）抗体免疫印迹试验（WB）确证不确定结果的血清学特点及临床意义,探讨导致电化学发光法筛查假阳性且免疫印迹试验（WB）确证不确定的临床原因。方法　对四川大学华西医院2013年HIV抗体WB确证结果为不确定的107例患者的带型及其临床资料进行分析,其中有WB随访结果的患者47例以WB随访结果进行判断,无WB随访结果的患者60例,结合核酸（HIV-1 RNA）或HIV-1 P24抗原定量测定结果进行综合判断。结果　107例WB确证不确定患者共有16种带型,p24比例最高,占51.4%,其次是gp160+p24,占15.89%。env类、pol类、gag类分别占28.97%、6.54%、64.49%。在env类中,阳性患者的出现率最高,为67.74%,pol类和gag类分别为14.29%、2.90%。16例HIV抗体ELISA法和电化学发光法同时筛查阳性的患者,其最终结果为阳性的发生率为93.75%。83例最终确证结果为阴性的患者中,肿瘤患者的构成比最高,占27.71%。结论　在HIV抗体WB确证不确定的结果当中,env类提示HIV感染的作用最大;gag类大部分为非特异性反应。ELISA法和电化学发光法同时阳性时提示HIV阳性的可能性较大。疾病因素与HIV抗体WB不确定的发生相关。     

HTLV-1 seroprevalence in aids patients and in HIV-1 seropositive and seronegative subjects at risk for aids in Northern Italy

Sera of 1023 subjects belonging to categories at risk for AIDS, positive for HIV-1, were tested for antibodies to human T-cell leukemia virus (HTLV-1) by ELISA, Western blotting, and radioimmunoprecipitation (RIP) assay. Sera of these subjects were positive for antibodies to HTLV-1 in a percentage of 18.54%. Nine out of 18 sera from patients with AIDS had antibodies to HTLV-1 (50%). Sera of 207 intravenous drug abusers and 64 homosexual males, seronegative for HIV-1, were tested for antibodies to HTLV-1 and 6.7% of intravenous drug abusers and 7.8% of homosexual males were found to be HTLV-1 antibody positive, showing that HTLV-1 infection occurred independently of HIV-1 and HIV-2 infections. In fact the same HIV-1 seronegative patients were also tested for the presence of antibodies to HIV-2 and none of them was found to be positive.Corresponding author.     

四种艾滋病病毒抗体筛查试剂检测性能评价

目的 通过对比分析艾滋病病毒(HIV)抗体筛查阳性结果与免疫印迹试验(WB)结果,评价4种HIV抗体筛查试剂检测性能.方法 2004年1月至2009年6月,分别用中山生物工程有限公司、荷兰生物梅里埃有限公司、珠海丽珠有限公司生产的3种酶联免疫吸附试验(enzyme linked immunasorbent assay,ELISA)试剂初筛血清HIV抗体,用美国雅培Determine HIV-1/2胶体硒标试剂、原试剂复检.筛查阳性样本用WB法进行确认.结果 共检测206 151例患者血清HIV抗体,确认HIV抗体阳性193例(0.094%).3种ELISA试剂敏感度、阴性预期值均为100%;雅培试剂分别为93.93%、91.67%,其漏检的样本均为WB不确定.中山、梅里埃、丽珠、雅培试剂的特异度分别为99.88%、99.89%、99.96%、89.38%;阳性预期值(study predictive value of a positive test result,PVP)分别为35.58%、46.46%、76.61%、92.20%;功效分别为99.88%、99.89%、99.96%、91.98%;3种ELISA试剂ROC曲线下面积分别为0.93、0.99、0.95.丽珠的PVP明显高于中山(X~2=45.804,P=0.000)、梅里埃(X~2=25.231,P=0.000);梅里埃的PVP比中山高,但无统计学意义(X~2=2.488,P=0.115);雅培PVP最高(与丽珠相比,X~2=18.633,P=0.000).在WB确认阳性、不确定、阴性组,均存在S/CO值[样本(sample)吸光度值/临界值(cut off)]＜6或≥6的样本.中山试剂确认阳性组S/CO值(14.29±2.63)明显高于阳性-阴性组(2.80±3.25)(t=17.652,P=0.000).梅里埃试剂确认阳性组S/CO值(16.09±2.35)明显高于阳性-阴性组(2.14±1.91)(t=31.622,P=0.000).丽珠试剂确认阳性组S/CO值(11.54±1.95)明显高于阳性-不确定组(5.54±3.57)(t=6.386,P=0.000)、阳性-阴性组(3.25±2.41)(t=21.772,P=0.000);阳性-不确定组S/CO值则高于阳性-阴性组(t=2.301,P=0.033).结论 4种筛查试剂性能良好,根据S/CO值不能准确估计WB确认结果,筛查阳性后必须进行确认.     

Monitoring for HIV-1, HIV-2, HTLV-I sero-progression and sero-conversion in a population at risk in east Africa.

Thirty-three individuals from East Africa, at risk for acquiring sexually transmitted infections, were selected to be monitored over a five month period for evidence of sero-progression and/or sero-conversion for human immunodeficiency virus type-1 and type-2 (HIV-1, HIV-2), and human T cell leukemia virus type-1 (HTLV-I). Initially, all sera were reactive by at least one retroviral screening assay, but most produced negative or indeterminate results by western blot assays. Five months after the initial screening, western blot assays indicated that one individual exhibited full sero-conversion for HIV-1; one HIV-1 positive individual also became positive for HIV-2; and two subjects showed sero-progression to become HTLV-I confirmed positive. Sera from fourteen individuals produced indeterminate results by western blot for HIV-1, ten of which were previously negative; the remaining four sera exhibited reactivity to at least one additional viral specific antigen after the five months. Circulating HIV-1 antigen was not demonstrated in any of the sera but DNA isolated from one of the individuals with indeterminate results produced a positive reaction for HIV-1 by the polymerase chain reaction.     

山东省羁押及收教人员HIV-1抗体血清学监测

目的：了解山东省监狱、劳教所羁押、收教人员艾滋病病毒（HIV）感染状况及特点,为制定防制措施提供依据.方法：采用EXCEL数据库和SPSS10.0软件对全省72 801名羁押、收教人员经筛查复检为HIV抗体阳性并经免疫印迹试验确认（WB）的结果及相关资料进行统计分析.结果：共发现HIV-1抗体阳性感染者21例,均为男性,感染率为0.0289%,其中外来人员13例,占总数的61.90%;HIV-1抗体不确定者9例,占确认数的28.13%.结论：我省羁押、收教人员HIV感染尚属低流行状态,但其为艾滋病传播的高危行为人群,应加强对该人群（尤其是外来男性）的监测与干预.     

2011-2014年常州地区HIV-1抗体阳性者 免疫印迹带型分析

摘要:目的　分析2011-2014年常州地区HIV-1抗体阳性者的免疫印迹（Western Blot,WB）带型。方法　应用WB试验对2011-2014年常州地区HIV-1抗体阳性待确证标本进行确证检测,并对其WB带型进行统计学分析。结果　1144份HIV-1抗体待确证标本中,36份确证为阴性,71份为不确定,1037份为阳性,阳性率为90.7%。在HIV-1抗体阳性标本WB带型中,gp160阳性率最高,为96.5%;p24和gp120次之,阳性率均在90.0%以上;gp41、p66、p51和p31阳性率较低,均低于90.0%;p17、p55和p39阳性率分别为66.4%、45.7%和41.9%;WB带型阳性率男女性之间差异均无统计学意义（P＞0.05）;20～39岁组的p39阳性率高于其他年龄组（χ2＝10.861,P＜0.05）,＜20岁组的p17阳性率高于其他年龄组（χ2＝7.906,P＜0.05）。结论　p39和p17带型阳性率在4个年龄组之间的差异均有统计学意义;gp160、gp120、gp41和p24是HIV-1抗体确证阳性的重要条带,且本地区检测的HIV-1抗体阳性者多在AIDS期前。     

2012 - 2017年武汉市HIV抗体确证检测结果分析

目的 对2012 - 2017年武汉市HIV初筛阳性样本的检测结果进行分析,了解武汉市HIV抗体确证情况。方法 依据《全国艾滋病检测技术规范（2009年版、2015年修订版）》对各筛查机构HIV筛查阳性反应的样本进行酶联免疫复核和免疫印迹确证试验。结果 2012 - 2017年武汉市艾滋病确证实验室共收到11 749份初筛阳性样本,经复核和确证试验,7 433份（63.26%）诊断为HIV - 1抗体阳性,3 780份（32.17%）为HIV - 1抗体阴性,536份（4.56%）为HIV - 1抗体不确定。阳性病例最多的年龄段为16 ～30岁。536例不确定样本中有156例（30.41%）进行了随访,120例（76.92%）确证为阳性,13例（8.33%）确证为阴性,23例（14.74%）仍为不确定。结论 2012 - 2017年武汉市HIV - 1抗体阳性数增长趋势明显,应进一步加强急性期和不确定结果病人的管理,做到早发现、早治疗。     

Antibodies to HTLV-1–2, HIV-1 and HIV-2 in syphilitic patients

Antibodies to human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) and to human T-cell leukemia virus (HTLV-1) were investigated by ELISA, Western blot and radioimmunoprecipitation (RIPA) assay in 318 sera (191 males and 127 females) obtained from syphilitic patients. The sera from 10% of the males and 3.1% of the females were positive for HIV-1. None of the sera contained antibodies to HIV-2. Antibodies to HTLV-1–2 were present in the sera of 7.1% of the males and 4.8% of the females who were seronegative for HIV. Five out of 24 (20.8%) HIV-1 positive subjects had antibodies to HTLV-1–2 as well.Sera from another group of 58 syphilitic patients (38 males and 20 females in the Anti-Venereal Disease Department), seronegative for HIV-1 and HIV-2, who denied both i.v. drug abuse and blood transfusion, were investigated in the same manner. None of the males had antibodies to HTLV-1–2, while 2 females (10%) were positive.     

孕产妇HIV抗体筛查阳性与免疫印迹试验对比

目的探讨孕产妇人类免疫缺陷病毒（HIV）抗体筛查试验阳性与免疫印迹试验（WB）结果的关系。方法对33份孕产妇HIV抗体待复查样本,进行酶联免疫吸附试验（ELISA）及免疫印迹试验（WB）,对WB试验结果不确定者进行3个月、6个月随访检测,6个月随访样本补充HIV-1病毒载量检测,分析ELISA检测与WB试验检测结果。结果 33份样本ELISA复检均呈阳性,免疫印迹试验（WB）中20份确认为HIV-1抗体阳性,ELISA检测与WB试验阳性符合率为60.61%（20/33）;S/CO值〉8的20份样本,WB试验结果均为HIV-1抗体阳性,1〈S/CO值〈8的13份样本,WB试验5份为HIV抗体阴性,8份为HIV抗体不确定;6个月随访样本HIV-1病毒载量检测,结果均小于最低检测限（50 IU/ml）。结论孕产妇HIV抗体筛查（ELISA）假阳性较高,假阳性样本存在于WB试验抗体阴性和不确定结果之中;随着ELISA试验S/CO值增高,ELISA检测与WB试验的阳性符合率随之增高。     

HIV抗体免疫印迹试验不确定结果的特征及鉴别方法研究

目的 研究HIV抗体确认不确定结果的血清学特征并比较不同实验方法鉴别不确定结果的效果.方法 以解放军艾滋病检测确认实验室2005-2006年确认的42例HIV抗体不确定者为研究对象,对血清样本进行HIV抗体条带免疫印迹检测、HIV病毒载量检测和HIV-1 p24抗原检测,同时进行随访,将随访前后的标本同时检测,比较免疫印迹带型的进展,判断HIV感染的真实状况,以随访的结果为标准,判断不同的方法鉴别不确定结果的效果.结果 (1)42例HIV抗体不确定病例中,共有8种带型:p24(45.2%)、gpl60(30.9%)、gp160±p24(11.9%).(2)对23例进行3个月以上随访,其中22例带型没有变化,判断为HIV抗体阴性;1例的带型在随访后出现显著进展,满足HIV抗体阳性诊断标准;随访证实,22例(95.6%)的不确定是非特异反应,只有1例是早期HIV感染.(3)对23例通过随访确定HIV感染状态的病例,条带免疫印迹检测1例血清学阳转的病例为阳性,22例血清学没有进展的病例中16例为阴性,6例仍为不确定,特异性是72.7%.HIV-1 p24抗原检测23份标本全部为阴性,1例血清学阳转标本漏检.检测7份血浆标本的病毒载量,1例血清学阳转的病例病毒载量为18 000 cp/ml,而6例血清学没有进展的病例检测结果全部低于最低检测限(＜LDL).结论 条带免疫印迹试验能够鉴别大约70.0%的不确定反应,病毒载量检测也是鉴别不确定结果的有效方法;使用这两种方法,可以对HIV抗体不确定进行早期鉴别诊断.     

Serological diagnosis of human immuno-deficiency virus in Burkina Faso: reliable, practical strategies using less expensive commercial test kits.

Reported are the results of a cross-sectional survey in Burkina Faso to identify reliable, practical strategies for the serological diagnosis of HIV-1 and/or HIV-2 infections, using less-expensive commercial test kits in various combinations, as an alternative to the conventional Western blot (WB) test, which costs US$ 60. Serum samples, collected from blood donors, patients with acquired immunodeficiency syndrome (AIDS) and pregnant women, were tested between December 1995 and January 1997. Twelve commercial test kits were available: five Mixt enzyme-linked immunosorbent assays (ELISA), three Mixt rapid tests, and four additional tests including monospecific HIV-1 and HIV-2 ELISA. The reference strategy utilized a combination of one ELISA or one rapid test with WB, and was conducted following WHO criteria. A total of 768 serum samples were tested; 35 were indeterminate and excluded from the analysis. Seroprevalence of HIV in the remaining 733 sera was found to be 37.5% (95% confidence interval: 34.0-41.1). All the ELISA tests showed 100% sensitivity, but their specificities ranged from 81.4% to 100%. GLA (Genelavia Mixt) had the highest positive delta value, while ICE HIV-1.0.2 (ICE) produced the most distinct negative results. Among the rapid tests, COM (CombAIDS-RS) achieved 100% sensitivity and SPO (HIV Spot) 100% specificity. Various combinations of commercial tests, according to recommended WHO strategies I, II, III, gave excellent results when ICE was included in the sequence. The best combination of tests for strategy II, which achieved 100% sensitivity and specificity, was to use ICE and COM, the cost of which was US$ 2.10, compared with US$ 55.60 for the corresponding conventional strategy. For strategy III, the best combination, which achieved 100% sensitivity and specificity, was to use ICE, ZYG (Enzygnost Anti HIV-1/HIV-2 Plus) and COM, the cost of which was US$ 2.90 (19.2 times lower than the corresponding strategy requiring WB). No rapid test combination showed 100% sensitivity and specificity. Our results indicate that the serodiagnosis of HIV in Burkina Faso is possible by using reliable, less-expensive strategies which do not require Western blot testing. Moreover, there is a choice of strategies for laboratories working with or without an ELISA chain.