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间接ELISA法检测轮状病毒

INDIRECT ELISA FOR DETECTION OF ROTA VIRUS

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【作者】 蒋慧惠严惠琴李洁宋广林滕庆杨铮刘纯荣陈晓蓝

【Author】 Jiang Hui-hui Department of Microbiology,School of Public Health,Shanghai First Medical College

【机构】 上海第一医学院卫生微生物学教研组上海第一医学院卫生微生物学教研组北京市儿童医院卫生部成都生物制品研究所

【摘要】 本文报道检测急性小儿腹泻患者粪便中轮状病毒的间接 ELISA 法。分别用兔抗 SA11轮状病毒血清和正常兔血清的粗提 IgG 包板,检测抗体为豚鼠抗 SA11抗体。酶标抗体为辣根过氧化物酶标记的抗豚鼠 IgG 抗体。根据正常人标本结果为阴性,阻断试验成立,以及阳性标本用正常豚鼠血清检测为阴性,说明本试验具特异性。分析比较50份标本的电镜检查结果和间接 ELISA 试验结果,说明本试验的敏感性好。初步比较了抗牛轮状病毒(NCDV)抗体和抗 SA11抗体检测的结果,两者基本一致。用硫酸铵粗提的 IgG 代替兔全血清,可减少非特异性显色。

【Abstract】 This paper reports the indirect ELISA for detection of rots virus in the stools of pati- ents with acute infantile diarrhea. The antibodies used were:(1)crude rabbit anti-SA-11 IgG for coating the wells;(2) guinea pig anti-SA-11 serum as the detecting antibody;and(3)HRP-anti guinea pig IgG as enzyme labelled antibody. The specificity of the test was proved by blocking test,normal stool specimens were negative,while possitive stool specimen showel negative results when normal guinea pig serum instead of guinea pig anti-SA-11 serum was used as the detecting antibody. The results of 50 stool specimens tested were comparable to that of the electron micros- copic examination,indicating that the sensitivity of the test was reliable. The anti-NCDV antiserum and antiSA-11 antiserum gave similar results. The crude rabbit anti-SA-11 IgG instead of whole rabbit antiserum was used,so that there was less nonspecificity.

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