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间接快速免疫组织化学试验检测多克隆抗 RNP IgG 抗体用于狂犬病诊断的评估。

Evaluation of polyclonal anti-RNP IgG antibody for rabies diagnosis by indirect rapid immunohistochemistry test.

机构信息

Instituto Pasteur, 393, Paulista Avenue, 01311-000, São Paulo, Brazil.

Instituto Butantan, São Paulo, Brazil.

出版信息

Acta Trop. 2020 Jun;206:105340. doi: 10.1016/j.actatropica.2020.105340. Epub 2020 Feb 21.

Abstract

Rabies still represents a major public health threat and estimated to cause 60,000 human deaths annually, particularly in developing countries. Thus, adequate surveillance based on rapid and reliable rabies diagnosis for both humans and animals is essential. The WHO and OIE recommended gold standard diagnostic technique for rabies is the direct immunofluorescence assay (dFAT). However, dFAT is expensive and requires a high level of expertise. As an alternative, the rapid immunohistochemistry technique is a promise to be a simple and cost effective diagnostic tool for rabies, and can be performed on field conditions prevalent in developing countries. However, no validated commercial conjugate antibody for rabies is available to meet the laboratory demand. Here, we evaluated the polyclonal anti-rabies virus ribonucleoprotein (RNP) IgG antibody for Rabies lyssavirus (RABV) detection by indirect rapid immunohistochemistry test (iRIT). We tested polyclonal anti-RNP IgG antibody against a batch of 100 brain specimens representing a wide phylogenetic origin in the State of São Paulo, Brazil. The purified IgG obtained 100% of diagnostic specificity and sensibility for RABV antigen detection in iRIT compared with the gold standard dFAT. In conclusion, our results demonstrate that the polyclonal anti-RNP IgG antibody may be used as a diagnostic reagent for rabies using iRIT, with the expectation of increase in availability and cost reduction of the epidemiological surveillance for developing countries.

摘要

狂犬病仍然是一个主要的公共卫生威胁,据估计每年导致 6 万人死亡,尤其是在发展中国家。因此,对人类和动物进行快速可靠的狂犬病诊断的充分监测至关重要。世界卫生组织和世界动物卫生组织推荐的狂犬病诊断金标准技术是直接免疫荧光检测(dFAT)。然而,dFAT 昂贵且需要高度专业知识。作为替代方案,快速免疫组织化学技术有望成为一种简单且具有成本效益的狂犬病诊断工具,并且可以在发展中国家常见的现场条件下进行。然而,目前没有经过验证的用于狂犬病的商业结合抗体来满足实验室需求。在这里,我们通过间接快速免疫组织化学试验(iRIT)评估了用于狂犬病弹状病毒(RABV)检测的多克隆抗狂犬病病毒核蛋白(RNP)IgG 抗体。我们针对来自巴西圣保罗州的代表广泛系统发育起源的 100 个脑标本批次测试了多克隆抗 RNP IgG 抗体。与金标准 dFAT 相比,纯化的 IgG 在 iRIT 中检测 RABV 抗原的诊断特异性和敏感性达到 100%。总之,我们的结果表明,多克隆抗 RNP IgG 抗体可用于 iRIT 检测狂犬病,有望增加发展中国家的流行病学监测的可用性和降低成本。

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