Development of a rapid immunochromatographic test for noroviruses genogroups I and II
Development of a rapid immunochromatographic test for noroviruses genogroups I and II
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DOI：
10.1016/j.jviromet.2007.10.010
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发表时间：
2008-03-01
影响因子：
3.1
通讯作者：
Ushijima, Hiroshi
中科院分区：
文献类型：
--
作者：
Takanashi, Sayaka;Okame, Michio;Ushijima, Hiroshi
已知诺沃克病毒(NOV)可导致全球儿童急性胃肠炎。虽然逆转录聚合酶链式反应(RT-PCR)方法被认为是诊断这种病毒感染的“金标准”，但它需要熟练的人员和设备齐全的实验室。本研究利用兔抗重组病毒样颗粒(RVLPs)的多克隆抗体，用免疫层析方法研制了一种快速、易操作的诊断试剂盒。该试剂盒对重组病毒样颗粒(RVLP)的反应性进行了评估，并与RT-PCR检测结果进行了比较。在前瞻性评价中，试剂盒的符合率为84.1%，敏感性为69.8%，特异性为93.7%。对RT-PCR阳性标本进行测序分型，发现部分异质性基因型也可检出，而同源基因型中的一些则偶尔出现假阴性，导致敏感性较低。未观察到与其他常见病毒病原体的交叉反应。结合病毒载量的检测下限为大约10(6-7)拷贝/克大便的结果，目前的免疫层析检测方法在简单的实验室支持下可以用于NOV感染的筛查。(C)2007 Elsevier B.V.保留所有权利。
Norovirus (NoV) is known to cause acute gastroenteritis in children worldwide. Although reverse transcription-PCR (RT-PCR) method is considered to be the "gold standard" for diagnosis of this viral infection, it requires skillful personnel and well-equipped laboratory. In this study, a rapid and easily performable diagnostic kit was developed using immunochromatographic method with rabbit polyclonal antibodies raised against recombinant virus-like particles (rVLPs) of most prevalent genotypes, genogroup II genotypes 3 and 4. This kit was evaluated for reactivity to rVLPs and detection of natural viruses in stool samples collected from children with diarrhea in comparison to the results obtained by RT-PCR. In the prospective assessment, the kit showed agreement rate of 84.1%, sensitivity of 69.8% and specificity of 93.7%. Genotyping of the RT-PCR positive samples by sequence analysis revealed that some heterogeneous genotypes were also detected while some in homogeneous genotypes occasionally showed false negative records resulting in lower sensitivity. No cross-reactivity with other common viral pathogens was observed. Taken together with the result of the detection limit of viral load as small as approximately 10(6-7) copies/g of stool, the current immunochromatography test is justified for screening for NoV infection with simple laboratory support. (C) 2007 Elsevier B.V. All rights reserved.