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.     

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。     

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。     

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.     

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.     

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。     

HIV抗体免疫印迹法检测中不确定结果的分析研究

目的 收集HIV抗体免疫印迹法检测中出现的不确定结果,统计分析各种带型的临床意义,为检验人员提供判定结果的方法和建议.方法 采集187份不确定实验结果和随访复检结果,以随访最终结果为金标准,统计分析各种带型出现的频度和阳性预测值,分析非特异性条带出现的频度和复检结果.结果 条带gp160、p24的阳性预测值分别为32....     

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。     

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.     

WHO international quality assessment scheme for HIV antibody testing: results from the second distribution of sera.

The WHO international quality assessment scheme for human immunodeficiency virus (HIV) antibody testing has been established to monitor the quality of laboratory performance in testing for antibodies to HIV. Following a small trial distribution of specimens early in 1989, the second distribution was made in February 1990. A total of 20 specimens of sera, 10 of which contained antibodies to HIV-1, were sent to 103 laboratories located in the six WHO Regions. Participants were asked to test the specimens using their routine methods and to report to WHO their findings on each specimen for each diagnostic assay used and their interpretation of the HIV antibody status of each specimen. For the antibody-positive specimens, 98.2% of the results were interpreted as positive and 1.8% as indeterminate; no false-negative interpretations were reported. For the antibody-negative specimens, 90.3% of the results were interpreted as negative, 1.3% as positive, and 8.4% as indeterminate. Most of the indeterminate reports were associated with one particular specimen. A wide variety of diagnostic assays and combinations of assays were used. In terms of the technical results obtained rather than their interpretation, the assays appeared extremely reliable for the positive specimens, with 99.5% of assay results being recorded as positive, 0.17% as negative, and 0.34% as indeterminate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Seroprevalence of HIV-1 and HIV-2 infection among children diagnosed with protein-calorie malnutrition in Nigeria.

Excessive weight loss due to protein calorie malnutrition (PCM) is a significant problem in Nigerian children. This syndrome may be difficult to differentiate from the wasting disease caused by human immunodeficiency virus type 1 (HIV-1) infection. We studied 70 children admitted to the Baptist Medical Center in Ogbomosho, Nigeria in 1990 with PCM for prevalence of antibodies to HIV-1 and HIV-2. The cohort was from low-risk mothers and had a median age of 25 months (range, 4 months-9 years) with a weight deficit of at least 20% of the theoretical weight for age. Two sera were positive for anti-HIV-1 by both ELISA and Western blot (WB). A high prevalence of samples negative for HIV-1 antibody by ELISA were repeatedly reactive (11%, 8/70) or indeterminate (46%, 32/70) by WB. None of the sera was positive for antibody to HIV-2. There was no correlation of ELISA positivity or extent of WB banding with successful recovery from malnutrition. These results indicate a relatively low but significant prevalence of HIV-1 infection in Nigerian children with PCM. The high prevalence of indeterminate reactions in WB assays for HIV-1 suggests that other procedures may be necessary for confirmatory diagnosis of HIV-1 infection in this African population.     

Evaluation of serological diagnostic indices for mucocutaneous leishmaniasis: immunofluorescence tests and enzyme-linked immunoassays for IgG, IgM and IgA antibodies

The sensitivity, specificity, positive predictive value, negative predictive value, and efficiency of immunofluorescence (IF) and enzyme-linked immunoassays (ELISA) for IgG, IgM and IgA antibodies were assessed on sera from mucocutaneous leishmaniasis patients and controls. The sensitivity of the IgG-ELISA test was 93.3% with 95% confidence interval higher than what could be due to a random test not associated with the disease. The specificity of all tests, except the IgM-ELISA, gave indices that could not have been due to chance. The IgG-ELISA and IgG-IF had the highest positive predictive value and the kappa statistic showed that the strength of agreement between the disease and the test was strongest for IgG-ELISA. The IgG-ELISA had a negative predictive value with 95% confidence limits that were not due to chance alone. Efficiency was highest for IgG-ELISA and IgG-IF. These results were obtained using sera from patients with severe or long-standing disease and from controls in whom the disease was ruled out by a negative Montenegro skin test. In field surveys where the differences between cases and controls are less easy to define the diagnostic indices of these tests may vary with the disease prevalence.     

Comparative evaluation of 36 commercial assays for detecting antibodies to HIV.

Summarized are the results of an assessment of the major operational characteristics of 36 commercially available assays for detection of antibodies to human immunodeficiency virus type 1 (HIV-1) and/or type 2 (HIV-2). For this purpose, 20 enzyme-linked immunosorbent assays (ELISAs), 11 simple immunoassays with visual reading, four supplemental assays, and one discriminatory assay were assessed using a panel of 537 sera (65% of which were of African, 26% of European, and 9% of South American origin); the prevalence of HIV-1 was 39.1% and of HIV-2, 15.7%. The following operational parameters of the assays were investigated: ease of performance; suitability for use in small blood collection centres; sensitivity and specificity; positive predictive values at different prevalences; inter-reader variability for simple assays whose results were read visually; the proportion of indeterminate results; and, for some of the ELISA assays, delta-values, as quantitative measures of sensitivity and specificity. The results will be of use to health policy decision-makers, managers of national AIDS prevention and control programmes, directors of blood banks, and laboratory specialists in the selection of appropriate HIV antibody assays.     

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。     

A seral epidemiological study of HIV transmitted through human seral gamma-globulin preparations.

In order to study the potential risk of transferring HIV through human seral gamma-globulin preparations (immunoglobulin), indirect immunofluorescent antibody test (IFA) and Western Blot (WB) assay were applied to 343 random samples (sera) with previous injection of imported human seral gamma-globulins (Ig) positive for Human Immunodeficiency Virus (HIV) antibodies between 1981-1987 for the detection of HIV antibodies. All results were negative and tests on all 23 controls who had previously received Ig made in China also gave negative results. However all 12 batches of imported Ig collected from the above-mentioned users, were positive for HIV antibodies when tested by WB and IFA. This study shows that under normal conditions, human seral gamma-globulin does not transmit HIV.     

广东省1991～1995年HIV－1抗体确认试验结果分析

１９９１年到１９９５年，广东省艾滋病监测检验中心共确认ＨＩＶ－１抗体初筛阳性及可疑样本１０３份，蛋白印迹（ＷＢ）试验结果１０３份样本中，阳性７０份（６７．９６％），可疑８份（７．７７％），阴性２５份（２４．２７％）。７０份阳性样本呈现的带型与阳性对照带型相同的有２９份，只缺Ｐ１５带的有１２份，只缺Ｐ１８、Ｐ１５带的有９份，其余的２０份缺失的带各不相同．但１００％的阳性样本都呈现ｇｐ１６０带，９７．１４％呈现Ｐ２４带，９８．５７％ＨＩＶ－１阳性样本ＷＢ带型中至少都有ｅｎｖ、ｇａｇ、ｐｏｌ带各一种，按我国诊断标准，结果容易判断．     

艾滋病筛查阳性标本复检与确认试验结果的对比分析

目的通过HIV抗体酶联免疫吸附（ELISA）初筛试验与蛋白印迹（WB）确认试验结果的比对,探讨目前艾滋病检测存在的问题。方法对芜湖市2011年艾滋病筛查阳性标本复检与确证实验结果进行比对,并分析WB确认试验带型及感染者基本信息。结果 119份HIV抗体初筛阳性血清经WB确认80份阳性,23份阴性,16份不确定。其中,两种初筛试剂测试结果均阳性的93份标本经WB确认80份阳性,7份阴性,6份不确定;两种初筛试剂测试结果一阴一阳的26份血样经WB确认16份阴性,10份不确定。在80份HIV-1确认阳性标本中,P55条带出现次数最少,仅为50.00%。在16份不确定标本中,Gp160条带出现频率最高,占总不确定标本的62.50%。结论筛查弱阳性的标本尽量用筛查试剂排除,从而减少＂HIV抗体不确定＂结果。不确定结果与WB试验的假阳性有关,可结合流行病学资料对结果进行准确解释。     

病毒载量检测有助于排除人免疫缺陷病毒抗体可疑结果中的非感染者

目的研究病毒载量检测是否能够辅助排除人免疫缺陷病毒（human immunodeficiency virus,HIV）抗体可疑样本中的非感染者。方法选择在筛查实验中两种酶联试剂检测结果不一致（一阴一阳、样本S／CO比值〈6）、且确认实验检测结果为“HIV抗体可疑”的样本19份测定病毒载量。对这些个体的感染状况进行跟踪随访,观察6个月后抗体进展情况。结果19份筛查实验结果不一致的抗体可疑样本的病毒载量检测均为阴性,随访复查条带无进展,均排除感染HIV的可能。结论在抗体筛查实验和确认实验反应均很弱的条件下,病毒载量检测阴性有助于排除非HIV感染原因造成的抗体检测中的可疑结果。     

南通市HIV抗体筛查阳性标本的确认结果分析

目的:通过对HIV抗体筛查(ELISA)阳性标本进行确认试验(WB),分析两者之间的关系。方法:按照《全国艾滋病检测技术规范》(2009版)的检测方法和要求,对南通市250例HIV抗体ELISA标本采用WB进行确认。结果:250例HIV抗体筛查阳性标本中,221例(88.4%)确认为HIV-1抗体阳性,22例(8.8%)为HIV抗体不确定,7例(2.8%)为HIV抗体阴性。阳性确认标本中,gp160、gp120、p66、p51、gp41、p31、p24的出现率均在90%以上,p55、p39和p17的出现率相对较低,分别为41.63%、37.56%、83.26%;不确定标本中,p24的出现率最高,为77.27%,其次是p160,为63.64%。结论:S/CO值越高,阳性确认率亦相应升高,但不能以此作为判别阳性的依据。对于不确定标本,除了加强随访外,还应积极寻求一种新的替代确认策略。     

四川省63例HIV抗体检测结果不确定者的追踪分析

目的分析造成HIV抗体不确定的可能因素,预测HIV抗体不确定者抗体进展情况。方法对HIV筛查阳性样本进行免疫印迹试验（WB）确认,对结果为HIV抗体不确定的检测者进行随访检测并对结果进行分析。结果不同人群,不同带型的检测者在进行复检时阳性率不同。临床期患者抗体滴度很低,确认为不确定时,病毒载量结果却很高。结论处于窗口期感染者、临床期患者以及处于某些特定时期的正常人群等都有可能导致HIV抗体检测出现不确定结果,必要时应结合其他检测方法,如病毒载量检测等,以及流行病学调查辅助做出准确的诊断。