使用单克隆抗体偶联红细胞检测和定量轮状病毒：与酶联免疫吸附测定法的比较

Detection and quantitation of rotavirus using monoclonal antibody coupled red blood cells: comparison with ELISA.

作者信息

Cranage M P, Campbell A D, Venters J L, Mawson S, Coombs R R, Flewett T H

出版信息

J Virol Methods. 1985 Aug;11(4):273-87. doi: 10.1016/0166-0934(85)90021-7.

Abstract

A total of 125 faecal extracts from infants were tested by reverse passive haemagglutination (RPH) using red cells coated with a monoclonal antibody against the major group-specific rotavirus antigen (VP 6). Results were compared with those obtained using a rabbit anti-rotavirus capture, guinea pig anti-rotavirus detector-based ELISA. The specificity of the assay was confirmed by use of 'normal' immunoglobulin coupled red cells and by inhibition with rabbit antiserum. The antibody-coated red cells could be stabilised by treatment with glutaraldehyde and subsequent freeze-drying with no detectable loss of activity even after storage at 45 degrees C for 4 wk. Good correlation was obtained between RPH and ELISA. Purified bovine rotavirus could be detected by RPH down to approximately 10(5) particles in a 25 microliters vol. Similar results were obtained with polyclonal antibody coupled cells and an ELISA using monoclonal antibody. Experiments using subgroup-specific monoclonal antibodies indicated the feasibility of rapid subgroup determination.

摘要

使用包被有抗主要群特异性轮状病毒抗原（VP6）单克隆抗体的红细胞，通过反向被动血凝试验（RPH）对125份婴儿粪便提取物进行了检测。将结果与使用兔抗轮状病毒捕获、豚鼠抗轮状病毒检测的酶联免疫吸附测定（ELISA）所获得的结果进行了比较。通过使用“正常”免疫球蛋白偶联红细胞以及用兔抗血清进行抑制，证实了该检测方法的特异性。用戊二醛处理后再进行冷冻干燥，可以使包被抗体的红细胞得以稳定，即使在45℃下储存4周后，其活性也没有可检测到的损失。RPH和ELISA之间具有良好的相关性。在25微升体积中，通过RPH能够检测到低至约10⁵个颗粒的纯化牛轮状病毒。使用多克隆抗体偶联细胞和使用单克隆抗体的ELISA也获得了类似结果。使用亚组特异性单克隆抗体的实验表明了快速进行亚组测定的可行性。