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Development of A Monoclonal Antibody-Coated Immunomagnetic Beads for Separation and Detection of Norovirus (Genogroup II) in Oysters

文献类型： 外文期刊

作者： Yao, Lin 1 ; Jiang, Yanhua 1 ; Jiang, Wei 1 ; Li, Fengling 1 ; Zhai, Yuxiu 1 ; Wang, Lianzhu 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Key Lab Test & Evaluat Qual & Safety Aquat Prod, Minist Agr, Qingdao 266071, Peoples R China

关键词： norovirus;oyster;immunomagnetic beads;separation;monoclonal antibody;RT-PCR

期刊名称：FOOD SCIENCE AND TECHNOLOGY RESEARCH （ 影响因子：0.668； 五年影响因子：0.895 ）

ISSN： 1344-6606

年卷期： 2013 年 19 卷 4 期

页码：

收录情况： SCI

摘要： The present study aimed to develop and evaluate a reverse transcription- polymerase chain reaction (RT-PCR) assay that incorporated immunomagnetic beads (IMS/RT-PCR) coated with a monoclonal antibody (Mab) for the separation and detection of noroviruses (NVs, genogroup II) in oysters. Mab-coated beads and polyclonal antibody (Pab)-coated beads were added to oyster extracts containing a series of tenfold dilution of NVs-positive fecal extracts. The oyster extracts were also treated with Proteinase K method. RNA was extracted and used as template for RT-PCR. Mab-based IMS/RT-PCR could detecte 10 RT PCR unit in oyster homogenate, it was 10 times more sensitive than Pab-based IMS/RT-PCR, and 102 times more sensitive than proteinase K/RT-PCR. Analysis of 473 oysters showed that Mab-based IMS/RT-PCR has the highest percentage of positive samples (5.29%), followed by Pab-based IMS/RT-PCR (3.81%), and proteinase K/RT-PCR (2.11%). Mab-based IMS/RT-PCR is a sensitive method for the detection of NVs in oysters and could be a good candidate for routine screening and risk assessment.

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