HIV的实验室检测技术进展

对目前实验室常用的艾滋病毒检测技术的前沿研究进展进行了综述,介绍了本实验室一种新的结合胶体金的H IV检测方法。     

HIV抗体检测技术研究进展

获得性免疫缺陷综合征,简称艾滋病(acquired immun-odeficiency syndrome,AIDS),是由免疫缺陷病毒(human im-munodeficiency virus,HIV)感染所引起的一种严重的传染性疾病。HIV/AIDS的流行已成为人类、社会和经济发展的灾难,全球活着的HIV感染者已超过4 300万人,每天约有14 000     

免疫渗滤法检测血液及尿液HIV抗体的应用评价

评价免疫渗滤法人类免疫缺陷病毒1+2型抗体诊断试剂盒检测人血浆和尿液样本的临床性能。采用对照试验研究,选取背景清晰的研究对象200例,采集同一研究对象的血浆和尿液样本,应用万泰生物药业公司生产的人类免疫缺陷病毒1+2型抗体诊断试剂盒作为考核试剂,法国生物梅里埃公司生产的人类免疫缺陷病毒抗体诊断试剂盒(ELISA法)作为参考试剂进行检测,考核试剂检测结果与参考试剂及研究对象背景进行比较分析。考核试剂检测血浆HIV抗体与参考试剂相比较,阳性符合率100.00%,阴性符合率100.00%,总符合率100.00%,Kappa值1.00,一致性强度为最强;考核试剂检测尿液HIV抗体与参考试剂检测结果相比较,阳性符合率68.29%,阴性符合率100.00%,总符合率87.00%,Kappa值0.72,一致性强度为高度。免疫渗滤法人类免疫缺陷病毒1+2型抗体诊断试剂盒对血浆、尿液样本检测性能优越,适合临床快速诊断。     

衣壳蛋白:抗HIV感染的新靶点

HIV是由病毒衣壳蛋白（capsid protein,CA）形成的一个蛋白质外壳包绕着内部的核蛋白复合体所构成。HIV-1的CA在病毒的组装和成熟中起着至关重要的作用。同时,病毒传染性很大程度上取决于衣壳的结构和稳定性,因此,衣壳蛋白成为潜在的抗HIV药物研究的重要靶点之一。     

HIV感染早期病毒p24蛋白的检测

目的:建立敏感、特异的检测血清中HIV-p24蛋白的方法,作为HIV感染早期即窗口期的监测手段。方法:用纯化的p24蛋白免疫小鼠及家兔,获得单克隆及多克隆抗体,经DEAE-52阴离子交换柱纯化后,标记辣根过氧化物酶,建立ELISA双抗体夹心及间接双抗体夹心方法,检测HIV-p24蛋白。结果:包被单抗、标记多抗或包被多抗、标记单抗,均能特异地检出系列稀释的p24蛋白,包被混合单抗较包被多抗更敏感;经标记的抗种属抗体放大可明显提高检测的敏感性。结论:建立了敏感、特异的检测p24蛋白的双抗体夹心法,间接放大方法可检出50pg/mL的HIV-p24蛋白,检测敏感性与国际同类产品相似。     

HIV的生物学特性

爱滋病(AIDS)目前正在全球蔓延,它已遍及五大洲,波及一百多个国家和地区。世界卫生组织估计,到1991年全世界将有50-300万AIDS患者,近1亿人受到HIV的感染,将成为世界十大致命疾病之一。本文就HIV的形态结构、基因组成、复制繁殖等生物学特性作简单的介绍: 一、概述 HIV是60年代分布在中非(扎伊尔、卢旺达、布隆迪、乌干达、坦桑尼亚、肯尼亚等国)绿猴身上的病毒变异而来的。70年代初期由维多利亚湖西部经中非传播,最早传到加勒比地区的海地,随后又传到美国,目前世界上AIDS患者中80%是美国人。1983年5月巴黎巴斯德研究院以蒙塔尼为首的科     

Dot-ELISA法检测血液及唾液HIV抗体的应用评价

评价人类免疫缺陷病毒1+2型抗体检测试剂盒(Dot-ELISA法)检测血清和唾液样本的临床性能。采用对照试验研究,选取背景清晰的研究对象200例,采集同一研究对象的血清和唾液样本,应用万泰生物药业公司生产的人类免疫缺陷病毒1+2型抗体检测试剂盒作为考核试剂,法国生物梅里埃公司生产的人类免疫缺陷病毒抗体诊断试剂盒(ELISA法)作为参考试剂,考核试剂检测结果与参考试剂及研究对象背景进行比较分析。考核试剂检测血清HIV抗体与参考试剂相比较,阳性符合率100%,阴性符合率100%,总符合率100%,Kappa值1.00,一致性为最强;考核试剂检测唾液HIV抗体与参考试剂检测结果相比较,阳性符合率98.78%,阴性符合率100%,总符合率99.50%,Kappa值0.99,一致性为最强。Dot-ELISA法人类免疫缺陷病毒1+2型抗体检测试剂盒对血清及唾液样本检测性能优越,适合HIV抗体快速筛查。     

HIV储存库特征及其主要检测方法

获得性免疫缺陷综合症(acquired immunodeficiency syndrome,AIDS),即艾滋病,是由人类免疫缺陷病毒(human immunodeficiency virus,HIV)引起的一种致死率极高的慢性传染病。如何有效控制和预防AIDS已成为全球共同面临的严峻挑战。HIV DNA整合在CD4+T细胞基因组中形成HIV储存库,使部分HIV逃避了宿主免疫清除和抗病毒药物的治疗作用。如何彻底清除患者体内HIV储存库成为功能性治愈艾滋病的重大障碍。了解HIV储存库的病毒学特征及其检测方法,是目前HIV研究的热点之一。现对HIV储存库特征及其主要检测方法作一综述。     

HIV感染检测技术的研究进展

ＨＩＶ感染的检测是防治艾滋病和控制其传播的重要手段。本文综述了从宿主标本中直接检测到病毒本身的ＰＣＲ技术的抗原检测,以及抗ＨＩＶ检测技术的应用情况及优缺点,并结合自己的工作实际对ＨＩＶ感染检测技术的选择使用进行评述     

HIV感染与硒衰竭

近年来硒在艾滋病发生发展中的作用越来越引起人们的关注,ＨＩＶ感染者及ＡＩＤＳ病人具有渐进性硒衰竭的症状,而病毒及Ｔ细胞有可能具有编码硒蛋白的功能,其中一种病毒编码的硒蛋白可能对ＨＩＶ的表达起抑制作用。在体外,硒对ＨＩＶ １的复制具有调节作用,为硒的临床应用提供了潜在的前景。     

Circulating microRNAs as Biomarkers for Detection of Autologous Blood Transfusion

MicroRNAs (miRNAs) are small non-coding RNAs that regulate various biological processes. Cell-free miRNAs measured in blood plasma have emerged as specific and sensitive markers of physiological processes and disease. In this study, we investigated whether circulating miRNAs can serve as biomarkers for the detection of autologous blood transfusion, a major doping technique that is still undetectable. Plasma miRNA levels were analyzed using high-throughput quantitative real-time PCR. Plasma samples were obtained before and at several time points after autologous blood transfusion (blood bag storage time 42 days) in 10 healthy subjects and 10 controls without transfusion. Other serum markers of erythropoiesis were determined in the same samples. Our results revealed a distinct change in the pattern of circulating miRNAs. Ten miRNAs were upregulated in transfusion samples compared with control samples. Among these, miR-30b, miR-30c, and miR-26b increased significantly and showed a 3.9-, 4.0-, and 3.0-fold change, respectively. The origin of these miRNAs was related to pulmonary and liver tissues. Erythropoietin (EPO) concentration decreased after blood reinfusion. A combination of miRNAs and EPO measurement in a mathematical model enhanced the efficiency of autologous transfusion detection through miRNA analysis. Therefore, our results lay the foundation for the development of miRNAs as novel blood-based biomarkers to detect autologous transfusion.     

Arginase Activity in the Blood of Patients with Visceral Leishmaniasis and HIV Infection

Background

Visceral leishmaniasis is a parasitic disease associated with high mortality. The most important foci of visceral leishmaniasis in Ethiopia are in the Northwest and are predominantly associated with high rates of HIV co-infection. Co-infection of visceral leishmaniasis patients with HIV results in higher mortality, treatment failure and relapse. We have previously shown that arginase, an enzyme associated with immunosuppression, was increased in patients with visceral leishmaniasis and in HIV seropositive patients; further our results showed that high arginase activity is a marker of disease severity. Here, we tested the hypothesis that increased arginase activities associated with visceral leishmaniasis and HIV infections synergize in patients co-infected with both pathogens.

Methodology/Principal Findings

We recruited a cohort of patients with visceral leishmaniasis and a cohort of patients with visceral leishmaniasis and HIV infection from Gondar, Northwest Ethiopia, and recorded and compared their clinical data. Further, we measured the levels of arginase activity in the blood of these patients and identified the phenotype of arginase-expressing cells. Our results show that CD4⁺ T cell counts were significantly lower and the parasite load in the spleen was significantly higher in co-infected patients. Moreover, our results demonstrate that arginase activity was significantly higher in peripheral blood mononuclear cells and plasma of co-infected patients. Finally, we identified the cells-expressing arginase in the PBMCs as low-density granulocytes.

Conclusion

Our results suggest that increased arginase might contribute to the poor disease outcome characteristic of patients with visceral leishmaniasis and HIV co-infection.