Comparison of enzyme-linked immunosorbent assay, electron microscopy, and reversed passive haemagglutination for detection of human rotavirus in stool specimens.

Biken journal Pub Date : 1983-06-01
H Tsuchie, K Shimase, I Tamura, O Kurimura, E Kaneto, T Katsumoto, M Ito, T Kurimura
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Abstract

An enzyme-linked immunosorbent assay (ELISA) using microplates as solid phase, rabbit antiserum against human rotavirus Wa strain as catching antibody, and the same reagent labeled with beta-D-galactosidase as conjugate, has been developed for detection of human rotavirus antigen(s) in stool specimens from patients with acute gastroenteritis. The limit of detection of purified human rotavirus by ELISA was 15.6 ng/ml (1.56 ng/well) of viral protein. The sensitivities of ELISA, electron microscopy, and the reversed passive haemagglutination method (ROTA-CELL) were compared. ELISA was more sensitive than electron microscopy and the reversed passive haemagglutination method. The ELISA blocking assay was useful for detection of an antibody response to human rotavirus in paired sera from children in two institutions during outbreaks of rotavirus gastroenteritis.

酶联免疫吸附法、电子显微镜和反向被动血凝检测粪便标本中人轮状病毒的比较
建立了一种酶联免疫吸附试验(ELISA),以微孔板为固相,兔抗人轮状病毒Wa株抗血清为捕获抗体,以β - d -半乳糖苷酶标记的试剂为偶联物,用于检测急性胃肠炎患者粪便标本中的人轮状病毒抗原。ELISA法对纯化的人轮状病毒蛋白的检出限为15.6 ng/ml (1.56 ng/孔)。比较ELISA法、电镜法和反向被动血凝法(ROTA-CELL)的敏感性。ELISA检测结果比电镜和反向被动血凝法更灵敏。在轮状病毒肠胃炎暴发期间,ELISA阻断试验可用于检测来自两个机构的儿童配对血清中对人轮状病毒的抗体反应。
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