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抗人嗜T淋巴细胞病毒1型p19单克隆抗体的制备与鉴定:用于特异性捕获酶免疫测定

Production and characterization of a monoclonal antibody directed against HTLV-1 p19: use in a specific capture enzyme immunoassay.

作者信息

Ebersold A, Noraz N, Grange J, Gasmi M, Grange M P, Souche S, Mamoun R, Desgranges C

机构信息

Unité de Recherche sur les Hépatites, le SIDA et les rétrovirus humains, INSERM U271, Lyon, France.

出版信息

Hybridoma. 1993 Apr;12(2):185-95. doi: 10.1089/hyb.1993.12.185.

Abstract

An enzyme immunoassay (EIA) was developed for detection of Human T-cell Leukemia Virus antigen in culture supernatants and cell lysates. The assay used a mouse monoclonal antibody against HTLV-I p19 major core protein as capture antibody. It has a sensitivity of 1 microgram/ml of HTLV-I protein, 250 pg/ml of purified recombinant p19 and detected p19 in an 10(-2) diluted supernatant of MT2 infected cell and in a 100 MT2 cells lysate (10(6) cells taken at day 7 of culture). The assay enable us to discriminate between HTLV-I and HTLV-II antigens and is reproducibly negative for supernatants and cell lysates of uninfected cells and of HIV-1 infected cells. The assay was found to be more specific and 10 times more sensitive than the reverse transcriptase (RT) assay, and the EIA test became positive three days earlier than RT assay for the HTLV-I cell lines supernatants.

摘要

开发了一种酶免疫测定法(EIA),用于检测培养上清液和细胞裂解物中的人类T细胞白血病病毒抗原。该测定法使用针对HTLV-I p19主要核心蛋白的小鼠单克隆抗体作为捕获抗体。它对HTLV-I蛋白的灵敏度为1微克/毫升,对纯化的重组p19的灵敏度为250皮克/毫升,并且能在MT2感染细胞的10^(-2)稀释上清液和100个MT2细胞裂解物(培养第7天取10^6个细胞)中检测到p19。该测定法使我们能够区分HTLV-I和HTLV-II抗原,对于未感染细胞以及HIV-1感染细胞的上清液和细胞裂解物,该测定法可重复性地呈阴性。发现该测定法比逆转录酶(RT)测定法更具特异性,灵敏度高10倍,并且对于HTLV-I细胞系上清液,EIA测试比RT测定法早三天呈阳性。

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