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通过酶联免疫吸附测定法检测火鸡肠道冠状病毒并与其他冠状病毒进行区分。

Detection of turkey enteric coronavirus by enzyme-linked immunosorbent assay and differentiation from other coronaviruses.

作者信息

Dea S, Tijssen P

机构信息

Department of Comparative Medicine, University of Quebec, Laval-des-Rapides, Canada.

出版信息

Am J Vet Res. 1989 Feb;50(2):226-31.

PMID:2541641
Abstract

A double-antibody ELISA for the detection of coronaviruses in intestinal contents from turkey poults with diarrhea was developed. Antibodies were raised in rabbits and guinea pigs against a Minnesota isolate of turkey enteric coronavirus (TCV) propagated in embryonating turkey eggs and were purified by density-gradient centrifugation. The specificity of antisera was confirmed by hemagglutination-inhibition and immunoelectron microscopy. Absorption of anti-TCV hyperimmune sera with egg extracts or egg ovalbumin and the use of different dilution and blocking buffers influenced the sensitivity and specificity of the ELISA. Reciprocal cross-reactivity was detected among turkey, chicken, bovine, and murine coronaviruses. Antisera to the transmissible gastroenteritis virus of swine, the rabbit enteric coronavirus, or the human coronavirus strain 299E failed to react with TCV. The TCV cross-reacted only moderately with the avian infectious bronchitis virus and the hemagglutinating encephalomyelitis virus of swine. Investigations with samples from 47 commercial turkey flocks in Quebec with episodes of transmissible enteritis revealed that the ELISA was more sensitive than was electron microscopy for detection of TCV.

摘要

开发了一种双抗体ELISA,用于检测腹泻火鸡雏鸡肠道内容物中的冠状病毒。用在鸡胚中繁殖的明尼苏达火鸡肠道冠状病毒(TCV)毒株免疫家兔和豚鼠,制备抗体,并通过密度梯度离心法纯化。通过血凝抑制试验和免疫电子显微镜证实了抗血清的特异性。用卵提取物或卵清蛋白吸收抗TCV超免疫血清,以及使用不同的稀释和封闭缓冲液会影响ELISA的敏感性和特异性。在火鸡、鸡、牛和鼠冠状病毒之间检测到交叉反应。抗猪传染性胃肠炎病毒、兔肠道冠状病毒或人冠状病毒299E毒株的抗血清与TCV无反应。TCV仅与禽传染性支气管炎病毒和猪血凝性脑脊髓炎病毒有中度交叉反应。对魁北克47个有传染性肠炎疫情的商业火鸡群的样本进行调查发现,ELISA检测TCV比电子显微镜更敏感。

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