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胶乳凝集试验检测诺如病毒方法的建立。

Development of a latex agglutination test for norovirus detection.

机构信息

Department of Environmental Health, Institute for Health and Environment, School of Public Health, Seoul National University, Seoul, Republic of Korea.

出版信息

J Microbiol. 2010 Aug;48(4):419-25. doi: 10.1007/s12275-010-0071-4. Epub 2010 Aug 20.

Abstract

Norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Currently, reverse transcription polymerase chain reaction (RT-PCR) is used commonly to detect NoVs in both clinical and environmental samples. However, RT-PCR requires expensive equipment and cannot be performed on site. In this study, a latex agglutination test (LAT) using antibody-labeled latex beads for detecting NoVs was developed. Two kinds of polyclonal antibodies, one generated from synthetic peptides and the other from E. coli-expressed NoV capsid proteins, were used to develop the LAT. Each of these polyclonal antibodies was immobilized on the surface of latex beads and tested for the ability to detect NoVs. Under optimized conditions, our LAT detected GII.4 NoV at concentrations as low as 3.3x10(5) RT-PCR units/ml in stool samples. The detection limit for the LAT was approximately 1.7 103 RT-PCR units. Forty-eight stool samples were tested for NoVs using this LAT. In comparison with an RT-PCR assay, the sensitivity and specificity of the LAT were 35% and 100%, respectively. With further optimization, this LAT used with appropriate antibodies could be applied for convenient detection of NoVs in clinical diagnosis and food monitoring.

摘要

诺如病毒(NoV)是全球急性胃肠炎的主要原因。目前,逆转录聚合酶链反应(RT-PCR)常用于检测临床和环境样本中的 NoV。然而,RT-PCR 需要昂贵的设备,并且不能在现场进行。在本研究中,开发了一种使用抗体标记乳胶珠检测 NoV 的乳胶凝集试验(LAT)。两种多克隆抗体,一种来自合成肽,另一种来自大肠杆菌表达的 NoV 衣壳蛋白,用于开发 LAT。这两种多克隆抗体都固定在乳胶珠的表面,并测试其检测 NoV 的能力。在优化条件下,我们的 LAT 在粪便样本中检测到浓度低至 3.3x10(5) RT-PCR 单位/ml 的 GII.4 NoV。LAT 的检测限约为 1.7x103 RT-PCR 单位。使用该 LAT 检测了 48 份粪便样本中的 NoV。与 RT-PCR 检测相比,LAT 的灵敏度和特异性分别为 35%和 100%。通过进一步优化,这种与适当抗体一起使用的 LAT 可以应用于临床诊断和食品监测中方便地检测 NoV。

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