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基于Env糖蛋白合成肽的猫免疫缺陷病毒感染的血清学诊断

Serological diagnosis of feline immunodeficiency virus infection based on synthetic peptides from Env glycoproteins.

作者信息

Avrameas A, Strosberg A D, Moraillon A, Sonigo P, Pancino G

机构信息

Institut Cochin de Génétique moléculaire, Unité d'Immunopharmacologie moléculaire et Génétique des Virus, CNRS UPR 415, France.

出版信息

Res Virol. 1993 May-Jun;144(3):209-18. doi: 10.1016/s0923-2516(06)80031-2.

Abstract

Feline immunodeficiency virus (FIV) is a lentivirus which infects domestic cats, causing an acquired immunodeficiency syndrome (AIDS). The aim of the present work was the development of an immunoassay for the diagnosis of FIV infection, using synthetic peptides from FIV envelope (Env) glycoproteins. Four peptides (8 to 11 amino acids long) corresponding to group-specific epitopes of FIV Env extracellular (SU) or transmembrane (TM) glycoproteins were synthesized. They were evaluated by enzyme-linked immunosorbent assay (ELISA) for immunoreactivity with sera from naturally or experimentally FIV-infected cats. One of these, P237, corresponds to a conserved nonapeptide of FIV TM, folded as a loop between two cysteines. ELISA performed with P237 on 171 sera from FIV-infected cats and 46 sera from specific-pathogen-free cats showed no false positive cases and 100% detection of infected cat sera. Moreover, 47 pet cat sera which were negative with a whole virus-based-ELISA were tested with the P237 ELISA: 2 out of 47 showed reactivity. FIV infection of these two cats was confirmed by radio-immunoprecipitation assay. Temporal studies performed on serial serum samples from experimentally infected cats detected antibodies to P237 three to five weeks after inoculation of virus. Thus, the P237 ELISA is a sensitive and specific immunoassay for early detection of antibodies to FIV. In addition, this synthetic nonapeptide is easier to produce and purify than virus preparations or recombinant proteins.

摘要

猫免疫缺陷病毒(FIV)是一种慢病毒,可感染家猫,引发获得性免疫缺陷综合征(AIDS)。本研究的目的是利用FIV包膜(Env)糖蛋白的合成肽开发一种用于诊断FIV感染的免疫测定法。合成了四种对应于FIV Env细胞外(SU)或跨膜(TM)糖蛋白组特异性表位的肽(8至11个氨基酸长)。通过酶联免疫吸附测定(ELISA)评估它们与自然感染或实验感染FIV的猫的血清的免疫反应性。其中一种,P237,对应于FIV TM的一个保守九肽,折叠成两个半胱氨酸之间的一个环。用P237对171份FIV感染猫的血清和46份无特定病原体猫的血清进行ELISA检测,未出现假阳性病例,感染猫血清的检测率为100%。此外,用基于全病毒的ELISA检测为阴性的47份宠物猫血清用P237 ELISA进行检测:47份中有2份显示出反应性。通过放射免疫沉淀测定证实了这两只猫感染了FIV。对实验感染猫的系列血清样本进行的时间研究在接种病毒后三到五周检测到针对P237的抗体。因此,P237 ELISA是一种用于早期检测FIV抗体的灵敏且特异的免疫测定法。此外,这种合成九肽比病毒制剂或重组蛋白更容易生产和纯化。

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