Aubin J T, Boulay D, Chapus A, Bréchot C, Laure F, Agut H
Laboratoire de Bactériologie-Virologie, CNRS EP 57, C.E.R.V.I., Hôpital Pitié-Salpêtrière, Paris.
Res Virol. 1995 Jan-Feb;146(1):75-9. doi: 10.1016/0923-2516(96)80592-9.
A new non-isotopic sandwich hybridization assay was developed to detect human immunodeficiency virus type 1 (HIV1) provirus amplified by the polymerase chain reaction. The sensitivity and specificity of this new technique using 96-well microplates as the support for the sandwich hybridization procedure and stable enzyme-linked oligomer as the detection probe were compared with those of Southern hybridization using a 32P-labelled oligomer probe. Three laboratories studied 437 peripheral blood mononuclear cell samples from 294 different subjects including both HIV1-seropositive and -seronegative individuals. The non-isotopic assay exhibited a sensitivity of 99.5% and a specificity of 99.1% when compared with the Southern procedure. In addition, the non-isotopic assay gives clear numeric data, is safe when handling, and is especially adapted to large-scale analyses.
开发了一种新的非同位素夹心杂交检测法，用于检测通过聚合酶链反应扩增的人类免疫缺陷病毒1型（HIV-1）前病毒。将这种以96孔微孔板为夹心杂交程序载体、以稳定的酶联寡聚物为检测探针的新技术的灵敏度和特异性，与使用32P标记寡聚物探针的Southern杂交法进行了比较。三个实验室研究了来自294名不同受试者的437份外周血单核细胞样本，这些受试者包括HIV-1血清阳性和血清阴性个体。与Southern杂交法相比，非同位素检测法的灵敏度为99.5%，特异性为99.1%。此外，非同位素检测法能给出清晰的数值数据，操作安全，特别适用于大规模分析。
