The Establishment and Application of Immunofluorescence Assay for Detection of Antibodies to HIV-1

Li-Jing-Yun; WANG Hong-Xia; BAO Zuo-Xi

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August 1997

Li-Jing-Yun, WANG Hong-Xia and BAO Zuo-Xi. The Establishment and Application of Immunofluorescence Assay for Detection of Antibodies to HIV-1[J]. Virologica Sinica, 1997, 12(4).

Citation: Li-Jing-Yun, WANG Hong-Xia, BAO Zuo-Xi. The Establishment and Application of Immunofluorescence Assay for Detection of Antibodies to HIV-1 .VIROLOGICA SINICA, 1997, 12(4) : 371.

免疫荧光检测HIV-1抗体方法的建立和应用

1.
北京军事医学科学院微生物流行病研究所

摘要

免疫荧光法检测HIV抗体是确认HIV感染的方法之一［l］，与蛋白印迹法相比，它的特异性更高，易于鉴别非特异反应，更为经济、快速、容易操作。为了拥有更多的HIV抗体检测手段，我们用感染和未感染HIV－l的HeLaCD“细胞制备抗原片，建立了免疫荧光检测HIV抗体的方法，并进行了初步应用。材料和方法1细胞和病毒HeLaCD4”细胞，引自美国BruceChesebro博士的实验室，用含10％热灭活小牛血清的RPMI1640培养基培养和传代。HIV－l株引自美国。2血清艾滋诊断试剂国家参比品，批号9602Z16份经蛋白印迹确认的HIV抗体阳性血清；10份健康人血清Z10份责疽及肝炎病人血清；月份HIV抗体初筛阳性血清。3制备抗原片Hel，。CD4”细胞长成单层后，加入05ml。滴度为10‘TCID50的HIV一三，吸附lh后加入新鲜培养基，37℃培养3d将感染的细胞用025％的胰酶消化、洗涤，稀释成浓度为5。104细胞／mL的细胞是液，同样处理一瓶未感染的细胞，将感染和未感染的细胞等量混合，15“滴入Ic孔镀膜载玻片上，37℃、5％Cq条件下培养8h细胞长成本层后，用冷丙酮固定10min，封片，置一20℃保存备用。4...
- 免疫荧光
- , HIV－1
- , HeLaCD_4~+细胞

The Establishment and Application of Immunofluorescence Assay for Detection of Antibodies to HIV-1

1.
Institute of Microbiology and Epidemiology

Abstract

Using the mixture of HIV -1 infected and no - infected HeLa CD4+ cells, the antigen slides were prepared and the immunofluorescence assay for detection of antibodies to human immunode-ficiency virus type 1 was established. Seventy - seven serum samples including the national stan-dard reference of HIV antibody reagents were tested. The sensitivity and specificity were 96. 4 %and 100%, respectively. Compared with the confirmation test of Western Blot, the coincident rate was 92. 6 %. The results indicated th...
- Immunofluorescence assay
- , HIV - 1
- , HeLa CD4~+ cells

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

The Establishment and Application of Immunofluorescence Assay for Detection of Antibodies to HIV-1

1. Institute of Microbiology and Epidemiology

Abstract: Using the mixture of HIV -1 infected and no - infected HeLa CD4+ cells, the antigen slides were prepared and the immunofluorescence assay for detection of antibodies to human immunode-ficiency virus type 1 was established. Seventy - seven serum samples including the national stan-dard reference of HIV antibody reagents were tested. The sensitivity and specificity were 96. 4 %and 100%, respectively. Compared with the confirmation test of Western Blot, the coincident rate was 92. 6 %. The results indicated th...