HIV抗体纳米免疫磁珠快速检测试剂的研制

目的 研制一种用纳米免疫磁珠层析技术用以检测HIV抗体的快速诊断试剂.方法 运用碳二亚（EDC）将重组的HIV抗原gp41、gp36偶联到200 nm的超顺磁纳米颗粒上,在硝酸纤维素（NC）膜上包被gp41、gp36抗原,制备成免疫层析检测卡,然后对检测卡进行性能分析评估.结果 对HIV抗体国家参考品血清盘（胶体金类）检测,符合要求;对20份HIV抗体阳性和600份抗体阴性临床血清检测,灵敏度为100%,特异性为98.5%.检测卡室温保存12个月性能稳定.结论 研制出一种纳米免疫磁珠HIV抗体检测试剂,具有检测简便、快速、稳定性好和适于现场检测等特点.     

HIV抗体纳米免疫磁珠快速检测试剂的研制

目的 研制一种用纳米免疫磁珠层析技术用以检测HIV抗体的快速诊断试剂.方法 运用碳二亚(EDC)将重组的HIV抗原gp41、gp36偶联到200 nm的超顺磁纳米颗粒上,在硝酸纤维素(NC)膜上包被gp41、gp36抗原,制备成免疫层析检测卡,然后对检测卡进行性能分析评估.结果 对HIV抗体国家参考品血清盘(胶体金类)检测,符合要求;对20份HIV抗体阳性和600份抗体阴性临床血清检测,灵敏度为100%,特异性为98.5%.检测卡室温保存12个月性能稳定.结论 研制出一种纳米免疫磁珠HIV抗体检测试剂,具有检测简便、快速、稳定性好和适于现场检测等特点.     

HIV抗体纳米免疫磁珠快速检测试剂的研制

目的 研制一种用纳米免疫磁珠层析技术用以检测HIV抗体的快速诊断试剂.方法 运用碳二亚(EDC)将重组的HIV抗原gp41、gp36偶联到200 nm的超顺磁纳米颗粒上,在硝酸纤维素(NC)膜上包被gp41、gp36抗原,制备成免疫层析检测卡,然后对检测卡进行性能分析评估.结果 对HIV抗体国家参考品血清盘(胶体金类)检测,符合要求;对20份HIV抗体阳性和600份抗体阴性临床血清检测,灵敏度为100%,特异性为98.5%.检测卡室温保存12个月性能稳定.结论 研制出一种纳米免疫磁珠HIV抗体检测试剂,具有检测简便、快速、稳定性好和适于现场检测等特点.     

HCV抗体检测试剂对血站HCV抗体初筛试剂的评价

目的 建立对血站献血人员血液中HCV抗体初筛试剂的评价方法，为选择合适匹配的初筛试剂提供依据。方法 用新研制出的抗HCV不同基因功能区抗体检测试剂对两种国产试剂和一种进口试剂的特异性和灵敏度进行考核评价。结果 两种国产试剂之间的特异性和灵敏度均存在一定差异。且总体质量略差于进口试剂。结论 用新研制出的抗HCV不同基因功能区抗体检测试剂可对初筛试剂进行评价，选择合适的初筛试剂匹配对控制献血员的血液质量至关重要。     

HIV抗体酶联免疫诊断试剂检测不同基因型抗体的研究

目的 分析不同HIV抗体酶联免疫诊断试剂对检测HIV不同基因型抗体的情况。方法 对不同地区的20份HIV抗体阳性样品中HIV核酸进行扩增，对PCR产物进行测序并进行基因型别分析。用不同试剂对系列稀释的不同基因型样品进行检测。结果 20份样品均为HIV RNA阳性，其中9份样品为HIV B亚型，9份样品为：HIV C或BC重组，2份为HIVAE重组。不同试剂对HIV不同基因型抗体的检测灵敏度无明显差异。结论 我国主要的商业化HIV抗体诊断试剂产品检测不同基因型抗体的能力无明显差异。     

四种HIV抗体确证试剂盒用于HIV感染检测性能的比较研究

目的:比较4种HIV抗体确证试剂盒的检测性能,为临床使用试剂提供参考。方法:使用2种免疫印迹法(Western blot, WB)试剂、2种重组/线性免疫印迹法(recombinant immunoblot assay/line immune assay,RIBA/LIA)试剂平行检测185份样本,采用McNermar...     

梅毒螺旋体抗体快速试剂的实验室性能评价

目的 使用梅毒螺旋抗体快速试剂国家参考品以及商品化的血清转化盘和不同临床时期的梅毒样本对目前市面主要使用的TP抗体快速检测试剂进行性能评价。方法 选取34批次梅毒螺旋抗体快速试剂对梅毒螺旋抗体快速试剂国家参考品以及商品化的血清转化盘和51份不同临床时期梅毒感染样本进行检测,分析不同试剂的检测性能差异并依据检测结果进行评分。结果 34批次梅毒螺旋抗体快速试剂有5批次产品不符合梅毒螺旋抗体快速试剂国家参考品的性能要求,均为阳性参考品符合率项目;34批次梅毒螺旋抗体快速试剂检测血清转换盘阳转天数最大相差4周时间;评价梅毒螺旋抗体快速试剂对51份临床样本检测阳性率为84.3%～100%。来自14个厂家的16批次试剂评分结果为84.3～99.4分。结论 部分梅毒螺旋抗体快速试剂对早期梅毒感染样本的检出率较低,一方面需要选取合适的评价样本,另一方面需要加强试剂生产的工艺稳定性。     

甲型肝炎病毒IgM抗体快速检测试剂国家参考品的建立

目的 建立甲型肝炎病毒IgM抗体快速检测试剂国家参考品,制订其质量标准,用于该类试剂的质量评价。方法 通过对200多份单采浆站收集到的血浆进行初筛,筛选出16份备选样本。经多家实验室协助标定,分析标定结果,确定参考品的组成及其质量标准,并对参考品稳定性进行评估。结果 本参考品由10份阴性参考品、1份最低检出限参考品和重复性参考品组成。质量标准为10份阴性参考品符合率（-/-）应为10/10,最低检出限参考品要求1:40稀释时检测阳性,重复性参考品要求1:15稀释时连续检测10次,结果应均为阳性,且显色度均一。稀释血浆应均为HCV、HBV和HIV标志物阴性。参考品在4℃放置3、7 d和反复冻融2次条件下相对比较稳定。结论 成功建立甲型肝炎病毒IgM抗体快速检测试剂国家参考品,可用于该类试剂的质量控制及评价。     

浅谈HIV抗体初筛检测质量控制

随着市场经济的建立和物质文化生活水平的提高,人们的质量意识也在增强。质量不仅仅关系医院的生存和效益,而且也直接影响人类的健康和安全。作为医学的重要组成部分,检验医学的进步极大地改变了临床诊断手段,提高了治疗水平,在疾病的预防、诊断和治疗等方面发挥着越来越重要的     

HIV-1/2抗体和P24抗原联合检测试剂盒的研制与评价

目的研制HIV-1／2抗体和P24抗原联合检测酶免疫试剂盒并评价其实用性。方法联合使用基因工程HIVI／2型抗原和抗HIVP24单克隆抗体包被酶联反应板，以辣根过氧化物酶标记的HIVI／2型抗原和生物素化的兔抗HIVP24抗体作为标记物，研制了联合检测HIVI／2抗体和P24抗原的ELISA诊断试剂，并对其特异性、敏感性、稳定性等进行评价和临床考评。结果检测P24抗原质控品的灵敏度可达0．2ng／ml；与雅培公司试剂比较检测78份AIDS患者血清和85份正常人血清、对照检测中国药品生物制品检定所研制的HIV参比血清，特异度和灵敏度均为100％。临床考核检测12051份各种血清，灵敏度为100％（543／543），特异度为99．48％（11448／11508）。试剂在37℃放置6d后，试验结果无明显差异。结论本试剂盒具备特异度强、敏感度高、稳定性好、操作简便等优点，可以一步检出HIV特异性抗体和HIVP24抗原，缩短了HIV感染的检测窗口期，适用于HIV感染的实验室诊断和流行病学调查。     

Long-term highly suppressed HIV-infected children and adolescents with negative rapid HIV tests due to significant antibody loss

BackgroundRapid HIV test devices are widely used throughout the world and are important as diagnostic tools with relatively high sensitivity and specificity. Loss of HIV specific antibodies in late-stage AIDS patients has previously been reported in patients with advanced disease (i.e., AIDS).ObjectiveTo study rate of antibody loss that may lead to false negative HIV-antibodies results in children and adolescents who received long term antiretroviral (ARV) treatment with persistently undetectable viral loads.Study designFive FDA approved rapid HIV test kits including Trinity Uni-Gold Recombigen HIV-1, OraQuick Advance HIV-1/2, Reveal G3 HIV-1, Clearview STAT-PAK HIV-1/2, and Clearview COMPLETE HIV-1/2 were used to test 98 stored samples from 27 patients. Samples were tested at baseline and at least twice in 6–14 years post initiation of ARV treatment and full viral load suppression.ResultsOf the 403 tests, 43 (10.7%) were found to be false-negative using rapid HIV kits. Loss of positivity was correlated with decrease of HIV antibody titer.ConclusionsThere is a slow but persistent loss of HIV specific antibodies in highly suppressed HIV infected children and adolescents that may lead to false-negative results in rapid HIV antibody tests. The temporal loss of signal is dependent on the baseline level of antibodies and the type of HIV rapid test kit used.     

Evaluation of the Bio-Rad Multispot HIV-1/HIV-2 Rapid Test as an alternative to Western blot for confirmation of HIV infection

BackgroundIn the United States, a new HIV diagnostic algorithm has been proposed that uses an HIV-1/HIV-2 antibody differentiation immunoassay instead of Western blot or immunofluoresence for confirmatory testing.ObjectivesTo evaluate the Multispot HIV-1/HIV-2 Rapid Test (Multispot) as an alternative to Western blot analysis for confirmation of HIV infection.Study designA series of 205 serum and plasma specimens positive for HIV-1 or HIV-2 were used to compare the performance of Multispot to a standard HIV-1 Western blot. Positive samples included 63 specimens from patients > 18 months of age, 33 proficiency survey specimens, and 109 specimens from nine commercial seroconversion and performance panels. In addition, 63 specimens from 51 HIV-exposed, uninfected children ≤ 18 months of age in various stages of seroreversion and 192 HIV-negative samples were tested. Specimens were initially screened using a 4th generation HIV Ag/Ab Combo assay.ResultsMultispot readily discriminated between individuals with HIV-1 or HIV-2 infection and those who were uninfected. Of the 205 samples repeatedly reactive by the 4th generation screening assay, infection status was correctly confirmed by Multispot in 83.9% (172/205) compared to 68.8% (141/205) for Western blot. Multispot detected HIV-1 earlier in 27.6% of low-titer antibody specimens called indeterminate by Western blot, and effectively reduced the number of indeterminate results in seroreverting HIV-1 exposed, uninfected infants and for HIV-2 infections misinterpreted as indeterminate or positive by HIV-1 Western blot.ConclusionsMultispot offers speed and simplicity over Western blot and has an excellent performance for differentiation and confirmation of antibodies to HIV-1 and HIV-2.     

实时荧光环介导等温扩增检测 HIV-1 DNA反应体系的建立

目的建立一种快速、敏感度高的实时荧光环介导等温定量检测HIV-1 DNA的方法。方法根据HIV病毒基因序列选择6个保守区域设计4条环介导等温扩增（ LAMP）引物,建立环介导等温扩增体系,同时在体系中加入SYBR GreenⅠ荧光染料,并评价该方法的特异性和敏感度。结果成功合成实时荧光环介导等温定量检测HIV-1 DNA反应体系,检测200例HIV感染者外周淋巴细胞HIV-1 DNA,其中有195例阳性,并检测50例健康对照结果均阴性。其中有HIV-1 DNA定量结果显示最低含量为51拷贝/ml,最高含量为8．21×106拷贝/ml,平均值含量为5．78×105拷贝/ml,其中病毒含量比较集中在105数量级范围,共162例占83．08％。采用10倍稀释法发现该方法最低检出限为50拷贝/ml。对乙型肝炎病毒、疱疹病毒、丙型肝炎病毒进行交叉实验结果均为阴性。结论HIV感染者外周血中几乎都存在HIV-DNA,实时荧光环介导等温扩增是一种快速、敏感度高、特异性强的方法,适合各级医院的临床检测。     

Evaluation of supplemental testing with the Multispot HIV-1/HIV-2 Rapid Test and APTIMA HIV-1 RNA Qualitative Assay to resolve specimens with indeterminate or negative HIV-1 Western blots

BackgroundThe use of Western blot (WB) as a supplemental test after reactive sensitive initial assays can lead to inconclusive or misclassified HIV test results, delaying diagnosis.ObjectiveTo determine the proportion of specimens reactive by immunoassay (IA) but indeterminate or negative by WB that could be resolved by alternative supplemental tests recommended under a new HIV diagnostic testing algorithm.Study designRemnant HIV diagnostic specimens that were reactive on 3rd generation HIV-1/2 IA and either negative or indeterminate by HIV-1 WB from 11 health departments were tested with the Bio-Rad Multispot HIV-1/HIV-2 Rapid Test (Multispot) and the Gen-Probe APTIMA HIV-1 RNA Qualitative Assay (APTIMA).ResultsAccording to the new testing algorithm, 512 (89.8%) specimens were HIV-negative, 55 (9.6%) were HIV-1 positive (including 19 [3.3%] that were acute HIV-1 and 9 [1.6%] that were positive for HIV-1 by Multispot but APTIMA-negative), 2 (0.4%) were HIV-2 positive, and 1 (0.2%) was HIV-positive, type undifferentiated. 47 (21.4%) of the 220 WB-indeterminate and 8 (2.3%) of the 350 WB-negative specimens were HIV-1 positive.ConclusionApplying the new HIV diagnostic algorithm retrospectively to WB-negative and indeterminate specimens, the HIV infection status could be established for nearly all of the specimens. IA-reactive HIV-infected persons with WB-negative results had been previously misclassified as uninfected, and HIV diagnosis was delayed for those with WB-indeterminate specimens. These findings underscore the limitations of the WB to confirm HIV infection after reactive results from contemporary 3rd or 4th generation IAs that can detect HIV antibodies several weeks sooner than the WB.     

Field performance of the INSTI HIV-1/-2 antibody test in two South African antenatal clinics

This was a prospective study that assessed field performance of the INSTI HIV-1/-2 antibody test (INSTI test) in two antenatal clinics in South Africa (SA). INSTI test was evaluated against rapid tests used at these clinics, and pooled nucleic acid amplification testing (NAAT) performed for individuals with negative rapid tests. Three hundred and eighty-six pregnant women were enrolled; 334 (86.5%) with negative results on the screening rapid test, and 52 (13.5%; 95% confidence interval [CI]: 10.2–17.3%) with positive results on screening and confirmatory rapid tests. INSTI test yielded the same results as other rapid tests in all participants, thus showing a 100% sensitivity (95% CI: 93.2–100.0%) and specificity (95% CI: 98.9–100.0%). Pooled NAAT was performed for 290 participants who had negative rapid tests, and yielded negative results in all pools. These data show excellent field performance of the INSTI test, and highlight that this test can be implementedat SA clinics.     

福建省健康人群脊髓灰质炎抗体水平调查

目的 了解福建省实施口服脊髓灰质炎减毒活疫苗（OPV）常规免疫和强化免疫活动以来健康人群脊髓灰质炎抗体水平,为人群免疫策略、维持无脊灰状态提供科学依据.方法 应用微量细胞中和试验法对5个地区人群进行脊髓灰质炎抗体水平检测,建立数据库,SPSS统计软件进行统计分析.结果 共检测400名1～59岁人群的血清标本,脊灰Ⅰ、Ⅱ、Ⅲ型抗体阳性率分别为99.0%、99.3%、97.5%,几何平均滴度（GMT）Ⅰ、Ⅱ、Ⅲ型分别为1∶79.1、1∶31.2、1∶24.7;各型抗体GMT均随年龄增长呈下降趋势.结论 福建省通过OPV常规免疫和强化免疫活动后,健康人群对脊灰已形成免疫屏障.各型抗体GMT随年龄增长呈下降的趋势应引起重视.     

Multicenter evaluation of Xpert HIV-1 viral load assay for HIV quantification in China

New approaches to increase HIV-1 testing and HIV-1 viral load (VL) monitoring are needed for people living with HIV (PLHIV) in China. The Xpert HIV-1 VL assay was prequalified by the World Health Organization in 2017 but has not been evaluated in China. A multicenter evaluation was conducted to assess the accuracy of the Cepheid Xpert HIV-1 VL assay compared to the Abbott RealTime HIV-1 assay in China. Overall agreement was seen in 558 of 562 specimens (99.29%) with a κ value of 0.962. Pearson's coefficient between the two assays was 0.943. Analyzed by the Bland-Altman method, the mean bias was −0.54 log₁₀ copies/mL, and 94.05% results fell within the 95% confidence limit of agreement (−1.248 to 0.168 log₁₀ copies/mL). The coefficient of variation of the Cepheid Xpert HIV-1 VL assay ranged from 0.61% to 1.55%, as determined by testing eight positive plasma specimens with three different lots on different days. Due to its simplicity, random-access, rapid turnaround time, and accuracy, the Xpert HIV-1 VL assay can be used in local hospitals and clinics that bear the burden of identifying and treating HIV patients in China.