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首页> 外文期刊>Food microbiology >Detection of human norovirus in cherry tomatoes, blueberries and vegetable salad by using a receptor-binding capture and magnetic sequestration (RBCMS) method
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Detection of human norovirus in cherry tomatoes, blueberries and vegetable salad by using a receptor-binding capture and magnetic sequestration (RBCMS) method

机译：受体结合捕获和磁隔离（RBCMS）方法检测樱桃番茄，蓝莓和蔬菜沙拉中的人类诺如病毒

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摘要

In this study, we developed a sensitive receptor-binding capture and magnetic sequestration (RBCMS) method capable of concentrating human norovirus (HuNoV) from various food samples within few hours. We found that distilled water was suitable for the elution of HuNoV from inoculated tomatoes and blueberries, and glycine buffer improved the elution of HuNoV from inoculated salad. A significant improvement in post-extraction RNA yield was achieved by sequentially heat-releasing and column-extracting over either technique alone. The viral recovery of the RBCMS method was significantly higher than both the same-day PEG method (90 min PEG precipitation) and the two-day PEG method (overnight PEG precipitation) with a recovery rate of 8.75%, 1.03% and 5.40%, respectively. The detection limit of HuNoV by RBCMS method was significantly improved to 0.056 RTU. The estimated minimal concentration powers (MCPs) were 6.11,30.48, and 63.60-fold for the same-day PEG, two-day PEG, and RBCMS methods, respectively. RNase protection assay suggests that the viral genome was protected from RNase attack by remaining within the viral capsid. The signal detected by the RBCMS method might be more biologically relevant, as it requires both intact viral capsid to bind to HBGA receptors and the presence of viral genome to be amplified. Overall, the RBCMS method takes significantly less time than current PEG precipitation methods, recovers a higher yield of HuNoV from various food samples, and hence exhibits higher sensitivity.
机译：在这项研究中，我们开发了一种灵敏的受体结合捕获和磁螯合（RBCMS）方法，能够在数小时内从各种食品样品中浓缩人类诺如病毒（HuNoV）。我们发现蒸馏水适用于从接种番茄和蓝莓中洗脱HuNoV，甘氨酸缓冲液改善了从接种沙拉中洗脱HuNoV的情况。通过单独使用任一技术依次进行热释放和柱提取，可以显着提高提取后RNA的产量。 RBCMS方法的病毒回收率显着高于当天PEG方法（90分钟PEG沉淀）和两天PEG方法（过夜PEG沉淀），回收率分别为8.75％，1.03％和5.40％，分别。 RBCMS法检测HuNoV的检出限显着提高到0.056 RTU。对于当日PEG，两天PEG和RBCMS方法，估计的最小浓缩能力（MCP）分别为6.11、30.48和63.60倍。 RNase保护测定表明，病毒基因组通过保留在病毒衣壳中而免受RNase攻击。 RBCMS方法检测到的信号可能在生物学上更相关，因为它既需要完整的病毒衣壳与HBGA受体结合，又需要存在病毒基因组进行扩增。总体而言，RBCMS方法比目前的PEG沉淀方法花费的时间少得多，可从各种食品样品中回收更高的HuNoV产量，因此显示出更高的灵敏度。

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  • 来源
    《Food microbiology》 |2012年第2期|p.420-426|共7页
  • 作者

    Liangwen Pan; Qigang Zhang; Xiang Li; Peng Tian;

  • 作者单位

    Shanghai Entry-Exit Inspection and Quarantine Bureau, 1208 Minsheng Road, Shanghai 200135, China;

    Shanghai Entry-Exit Inspection and Quarantine Bureau, 1208 Minsheng Road, Shanghai 200135, China,School of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China;

    Shanghai Entry-Exit Inspection and Quarantine Bureau, 1208 Minsheng Road, Shanghai 200135, China;

    Produce Safety and Microbiology Research Unit, Western Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Albany, CA 94710, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    human norovirus (HuNoV); receptor-binding capture and magnetic; sequestration (RBCMS); quantitative real time RT-PCR (qRTPCR); virus recovery; encapsidated viral RNA;

    机译：人类诺如病毒（HuNoV）;受体结合捕获和磁性;隔离（RBCMS）;定量实时RT-PCR（qRTPCR）;病毒恢复;衣壳病毒RNA;

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