热灭活处理对HIV抗体检测影响的实验研究

用５６℃３０ｍｉｎ和６５℃１ｈ热灭活方法处理标本（包括ＨＩＶ抗体阳性的和正常的血清、血浆球蛋白的样品）,然后用ＥＬＩＳＡ、ＰＡ和ＷＢ检测ＨＩＶ抗体。其实验结果与一另一组未经处理的同份标本检测结果相同。说明标本热灭活的处理方法是一种简便有效的实验室安全防治措施,可以避免实验室ＨＩＶ感染的发生。     

样品热灭活对HIV抗体筛查和确证试验结果的影响研究

目的研究血浆样品经热灭活(56℃30min)后,其艾滋病病毒(HIV)抗体筛查和确证试验结果是否会受到影响。方法取5份HIV抗体阳性血浆样品,各分为2管,其中1管进行热灭活处理,然后对这2组样品分别进行10倍系列稀释,用1种HIV抗体酶联免疫吸附试验(ELISA)试剂检测,比较S/CO比值。取300份血浆样品,分为灭活组和普通组,分别用5种HIV抗体ELISA试剂(其中第三代试剂3种、第四代试剂2种)和3种HIV抗体快速检测试剂进行筛查检测,出现筛查阳性反应的样品进一步做确证试验。结果随着稀释度的增加,5份HIV抗体阳性样品灭活前后的ELISA检测结果(S/CO比值)都逐渐减小,直至转为阴性反应,其中4份样品灭活后比灭活前早一个稀释度转阴,1份同时转阴,提示灭活过程会略微降低HIV抗体的浓度。用5种ELISA试剂、3种快速检测试剂检测300份样品,样品灭活前后的检测结果差异均无统计学意义;对出现筛查阳性反应的所有样品,灭活前后的确证试验检测结果一致。结论经56℃30min热灭活后,尽管HIV阳性血浆样品中的HIV抗体浓度略有降低,但对于未经稀释的常规临床血浆样品来说,热灭活处理不会明显影响目前常用筛查、确证试剂的检测结果。     

70份HIV抗体初筛阳性结果分析

艾滋病病毒（HIV）抗体检测是目前诊断是否感染HIV的主要依据。六安市艾滋病筛查中心实验室承担本市监测哨点和本中心门诊的HIV抗体初筛实验，辖区各医疗机构HIV抗体筛查实验室初筛阳性标本的复判，以及将初筛阳性标本送省疾病预防控制中心（CDC）HIV确认实验室进行确认的工作。现将2000-2004年70份HIV抗体初筛阳性标本的检测结果分析如下。     

常温下邮政特快专递传送HIV抗体初筛阳性血清影响因素的研究

目的为安全、及时有效地传送艾滋病病毒（HIV）抗体初筛阳性血清,研究在常温下通过邮政特快专递途径传送HIV抗体初筛阳性血清的可行性,以确定温度、震荡、负压环境对HIV抗体阳性血清传送和检测的影响.方法选取不同S/CO值的HIV抗体阳性血清做震荡实验和热稳定实验.选用美国AXYGEN公司2.0ml冻存管（SCREW CAP TUBES SCT-200-SS-A）作为HIV抗体阳性血清的传送容器.用稀释墨水做负压实验和破坏实验.结果对于不同反应的HIV抗体阳性样本,在经过50℃加热88小时后,样本免疫印迹（WB）试验反应条带的结果没有任何差异;震荡不会影响HIV抗体检测;负压环境不会发生样本泄漏.常温下通过特快专递方法传送HIV抗体初筛阳性血清,符合《中华人民共和国邮政法》和《中华人民共和国邮政法实施细则》的规定.结论常温下通过邮政特快专递途径传送HIV抗体初筛阳性血清是可行的.     

我国现行HIV检测策略和方法的应用评价

目的评价我国现行HIV检测方法和程序的应用效果.方法按照我国<全国艾滋病检测工作规范>规定的方法和程序,分3个步骤对7 502份职业献血员的血清标本进行了HIV抗体检测,用一种国产试剂进行初筛,阳性标本用原有试剂和另外一种不同原理的试剂进行复核,两种试剂均阳性和一阴一阳的标本用免疫印迹试剂进行确认.结果初筛能够发现所有的HIV阳性,排除98.8%HIV阴性,但是有0.9%假阳性;复核实验能够在确认之前排除93.9%假阳性.确认实验最终确认19份标本HIV抗体阳性,4份标本HIV抗体可疑.90.9%假阳性和75.0%可疑标本的s/CO值在1.000～1.999之间,而94.7%阳性标本的s/CO值在5.000以上.结论复核实验对于有效排除假阳性,提高确认的效率,减少开支很有必要.s/CO值≥5.000预示阳性的价值比较大,s/CO值在1.000～1.999范围内多为假阳性或可疑.     

吸毒人群尿液血液标本HIV-1抗体ELISA检测对比分析

目的　用酶联免疫吸附试验 (ELISA)检测吸毒人群尿液标本中艾滋病病毒 1型 (HIV 1 )抗体 ,与血清学检测结果进行比较。方法　采集某市劳教所吸毒者尿液、血液标本 354例 ,HIV 1抗体阳性吸毒者复检尿液标本1 9例 ,共计 373例。应用ELISA初筛试剂检测尿液及血液标本中HIV 1抗体 ,阳性者取其血液标本进一步用Genelabs试剂做蛋白印迹 (WB)试验确认。结果　尿液、血液标本中检测HIV 1抗体的特异性为 99 72 % ,尿试剂的假阳性率为 0 2 8% ;血液标本HIV 1抗体阳性者 ,尿液标本检测也呈阳性 ,灵敏度为 1 0 0 %。结论　尿液标本用ELISA方法检测HIV 1抗体与血液标本ELISA方法的检出率有高度的一致性。尿试剂适用于对高危人群进行尿液HIV 1抗体的筛查工作     

一例HIV抗体蛋白免疫印迹法检测条带不全病例的分析

目的通过对最近一例艾滋病病毒(HIV)抗体实验室检测"不确定"结果的分析讨论,尽可能减少实验室"不确定"结果的比例,提高实验室HIV确证实验的质量。方法该病例的血清标本采用两种酶联免疫吸附试验(ELISA)及两种快诊实验检测,又用蛋白免疫印迹法(WB)进行确证实验,并结合流行病学资料、核酸检测和基因测序结果,对该样本进行全面分析。结果 2012年11月至2013年3月期间,一例首次来自上海市肺科医院,两次随访均来自浦东新区自愿咨询检测门诊(VCT)的病例,ELISA和快诊检测为HIV抗体阳性,但WB确证结果首次为"不确定",第2、第3次随访结果也均为"不确定",后经核酸检测结果为阳性,基因测序结果为HIV-1型的CRF01_AE重组亚型,符合该病例的流行病学调查资料。结论对确证试验为"不确定"的病例,应综合初筛结果和实验室辅助检测手段及流行病学史,尽早明确个体HIV感染情况。     

我国现行HIV检测策略和方法的应用评价

目的 评价我国现行HIV检测方法和程序的应用效果。方法 按照我国《全国艾滋病检测工作规范》规定的方法和程序,分3个步骤对7502份职业献血员的血清标本进行了HIV抗体检测,用一处国产试剂进行初筛,阳性标本用原有试剂和另外一处不同原理的试剂进行复核,两种试剂均阳性和一阴一阳的标本用免疫印迹试剂进行确认。结果 初筛能够发现所有的HIV阳性,排除98.8%HIV阴性,但是有0.9%假阳性;复核实验能够在确认之前排除93.9%假阳性。确认实验最终确认19份标本HIV抗体阳性,4份标本HIV抗体可疑。90.9%假阳性和75.0%可疑标本的s/CO值在1.000-1.9999之间,而94.7%阳性标本的s/CO值在5.000以上。结论 复核实验对于有效排除假阳性,提高确认的效率,减少开支很有必要。s/CO值≥5.000预示阳性的价值比较大,s/CO值在1.000-1.9999范围内多为假阳性或可疑。     

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。     

不同人群血清SARS病毒抗体及Ⅳ型胶原检测结果分析

为了解山东省传染性非典型肺炎 (SARS)病毒感染情况 ,以及 SARS患者是否出现肺部纤维化倾向 ,我们于 2 0 0 3年 6月 10日始采用血清学方法对临床诊断为 SARS的患者、与患者密切接触的医务人员及部分健康人的血清标本进行特异性SARS病毒抗体和 型胶原检测。现报告如下。1　材料与方法临床诊断为 SARS患者的血标本 1份 ,与该患者密切接触的医务人员血标本 33份 ,健康人血标本 10 2份。标本的采集均严格按照卫生部和 WHO制定的实验室生物安全手册执行。将血清经 5 6℃ 30分钟灭活后用于实验。用酶联免疫法检测 SARS病毒抗体。试剂…     

Hepatitis G virus in clotting factor concentrates

Blood-borne hepatitis is a well-known complication in patients with bleeding disorders. A recently discovered parentally transmitted virus, hepatitis G [GB virus C (GBV-C)] has an increased prevalence in patients with haemophilia. Clotting factor concentrates derived from pools of human plasma currently undergo viral inactivation techniques known to be effective against hepatitis B, C and HIV; however, the effectiveness of current purification and viral inactivation techniques against newly discovered viruses such as GBV-C is unknown. A total of 37 vials of clotting factor concentrates manufactured in the USA from 1981 to 1995 were tested for the presence of GBV-C virus. All samples that did not undergo a specific viral inactivation step were positive for GBV-C. Viral inactivation techniques that did not uniformly remove GBV-C included vapour heat treatment and dry heat treatments for less than 144 h. All samples treated by pasteurization, solvent detergent or dry heat for 144 h, were negative for the presence of GBV-C.     

Evaluation of the human immunodeficiency virus type 1 and 2 antibodies detection in dried whole blood spots (DBS) samples

Human Immunodeficiency Virus Type 1 and 2 antibodies detection was performed in 457 dried whole blood spots samples (S&S 903). Q-Preven HIV 1+2 was the screening test used. The results were compared with the gold standard serum tests by ELISA (Cobas Core e Axsym HIV1/2 gO) and immunofluorescence was the definitive confirmatory test. The samples were obtained from the Hospital Nossa Senhora da Concei??o in Porto Alegre, RS - Brazil, through whole blood transfer to filter paper card and sent to Caxias do Sul, RS-Brazil where the tests were performed. The dried whole blood spot stability was evaluated with two different panels. The first one was composed of five negative and five positive samples stored at room temperature, 4 degrees C, -20 degrees C and -70 degrees C, while the second was composed of two negative and three positive samples stored at 37 degrees C (humidity <50%). Each sample was screened every week for six weeks. These measurement results didn't show variation during the study period. The detected sensibility was 100%, specificity was 99.6%, the positive predictive value was 99.5% and negative predictive values were 100%. The results demonstrated high performance characteristics, opening a new perspective of dried whole blood spot utilization in HIV screening diagnosis.     

136份初筛抗-HIV1+2阳性而复检阴性标本的分析

目的了解山东省历年来艾滋病病毒(HIV)抗体检测中筛查试验假阳性结果的真实情况,更加合理地开展HIV抗体日常检测。方法检测及结果判断按《全国艾滋病检测技术规范》进行,对比分析HIV抗体筛查试验为阳性而免疫印迹试验(WB)为阴性的标本的结果以及相关资料。结果136份筛查试验为阳性反应、WB确认为HIV抗体阴性的标本,占所有送检标本的6.70%,占筛查试验阳性标本的13.39%。136份标本中,明胶颗粒凝集试验(PA)阳性的119份,占87.50%;酶联免疫吸附试验(ELISA)阳性33份,占24.26%,S/CO值平均为0.8005(0.2319~5.544)。结论日常检测必须遵从先筛查再确认的检测程序,筛查试验阳性的不能直接出HIV抗体阳性报告,应确认后再出。     

Loss of Red Blood Cell Viability Associated with Limited Thermal Inactivation of Extracellular HIV-1

The effects of incubation at mildly elevated temperatures on HIV-1 inactivation and in vitro red blood cell properties were investigated. Red cells (55% Hct) were leukodepleted (3 log₁₀) by filtration, maintained at 45 or 47°C for 4 or 8 h, and then stored at 4°C. Hemolysis was twice that of controls after 42-day storage for samples treated for 4 h at 45°C, and five times larger for samples heated at 47°C. There was also a significant increase in the rate of potassium loss, an early decrease in ATP levels, and an initial drop in pH for samples treated at either temperature. Larger differences were observed for samples exposed to these elevated temperatures for 8 h. Osmotic deformability curves obtained by ektacytometry showed dramatic decreases in red cell deformability at both temperatures and for both time periods. HIV-1 inactivation in red cells treated at 45°C (approximately 0.25 log₁₀/h) was considerably less than that obtained in tissue culture medium (1–2 log₁₀/h). Since the decrease in red cell deformability is likely to indicate reduced red cell function and survival, and the rate of HIV-1 inactivation is low, mild heat treatment is not an adequate process for viral inactivation of red cell products.     

Evaluation of an HIV screening assay kit for the combined detection of anti-HIV-1/2 antibodies and HIV p24 antigen]

We have evaluated a new HIV screening assay kit (Genscreen HIV Ag-Ab) for the HIV antigen-antibody combined test by comparing with two HIV antigen-antibody combined assay kits (VIDAS HIV DUO, Enzygnost HIV integral). Genscreen HIV Ag-Ab is a microwell plate enzyme immunoassay for the detection of HIV infection, based on the detection of anti-HIV-1/2 antibodies and HIV p24 antigen in human serum or plasma. In this study, 90 samples of HIV-1 antibody positive sera and 670 samples of HIV negative sera were examined. The sensitivity was 100% and the specificity was 99.7%. All of HIV-1 group M sera (subtypes A to G and B/D), HIV-1 group O sera and HIV-2 sera in worldwide HIV performance panel-302 were positive with Genscreen HIV Ag-Ab. Ten commercially available HIV-1 seroconversion panels were tested to evaluate sensitivity of three HIV antigen-antibody combined assay kits. Genscreen HIV Ag-Ab detected infection at the same bleeds as VIDAS HIV DUO in 8 of 10 seroconversion panels and 1 to 2 bleeds earlier than Enzygnost HIV integral in 5 of 10 seroconversion panels. However, VIDAS HIV DUO indicated false negative on 5th bleed in panel BB (PRA952). The result of the specimen was positive on 3rd bleed, equivocal on 4th bleed, negative on 5th bleed and again positive on 6th bleed. All of these specimens were positive by Genscreen HIV Ag-Ab. Therefore, Genscreen HIV Ag-Ab that shorten the window period is a useful and reliable for HIV screening test, especially in case of primary infection.     

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。     

Detection of HIV antibodies in saliva as a tool for epidemiological studies.

OBJECTIVE: To evaluate the use of saliva specimens for the detection of HIV antibodies among high-risk groups in epidemiological studies. DESIGN: Testing of saliva specimens collected by different methods from individuals with known HIV status. The most reliable method was examined for its usefulness in a field study among a high-risk group. METHODS: Saliva samples were obtained either by using a cotton-wool roll ('Salivette') or as 'whole saliva'. HIV antibodies were determined using commercial enzyme-linked immunosorbent assays (ELISA). Confirmation was performed using a line immunoassay or an immunoblot assay. RESULTS: In 'Salivette' samples, HIV antibodies were detected by ELISA in seven out of 22 seropositive individuals. In contrast, testing of 'whole saliva' samples from 79 HIV-seropositive and 115 HIV-seronegative individuals resulted in a 100% correlation with HIV serum status. The positive reaction of 20 'whole saliva' specimens was confirmed in a line immunoassay, whereas in an immunoblot assay only seven specimens were positive, one negative, and 12 indeterminate. In an HIV prevalence study among drug users, 395 'whole saliva' samples were tested in two different ELISA. Both assays showed complete agreement in detecting 58 positive and 337 negative samples. All positive samples were confirmed by the line immunoassay. CONCLUSION: Our study demonstrates that 'whole saliva' specimens are a good alternative to blood samples in epidemiological studies of HIV prevalence in high-risk groups.     

天津市MSM HIV感染窗口期检测策略的研究

目的探索适用于男男性行为人群（MSM）的艾滋病病毒（HIV）早期感染筛查策略,并对利用早期感染数据估测新发感染率的可行性进行评价。方法常规HIV抗体检测结合集合核酸检测,样品集合及分拆方法按50∶5∶1进行。结果 5 287份MSM血样中,5 073份HIV抗体检测阴性,5份样品为HIV抗体不确定,209份HIV抗体阳性。经核酸检测,5 073份HIV抗体阴性样品中,检出10份阳性;5份HIV抗体不确定样品均为阳性。15份核酸阳性样品中,有8例进行随访检测,均出现HIV抗体阳转。结论常规HIV抗体检测结合集合核酸检测可以发现更多的感染者,降低检测成本,对MSM中HIV感染窗口期诊断和HIV发病率的估测有重要意义。     

Syphilis serology and human immunodeficiency virus positivity in Chandigarh

AIDS and other sexually transmitted diseases are interlinked. VDRL positivity may indicate that the individual has an increased risk of being HIV positive as the epidemiological risk factors for developing syphilis and AIDS are similar. We analysed 323 (5.8%) VDRL positive serum samples (out of 5592 screened) for HIV positivity. All were HIV negative. While syphilitic infection in India at present is not commonly associated with HIV infection, experience in other countries indicates caution.