Human noroviruses impose a considerable health burden globally. Here, a flow cytometry approach designed for their detection in biological waste and food samples was developed using antibody-coated magnetic beads. Antipeptide antibodies against murine norovirus and various human norovirus genotypes were generated for capture and coated onto magnetic beads. A flow cytometry assay was then implemented to detect bead-bound human norovirus GI.3 in patient stool samples and in norovirus-spiked mussel digestive tissues. The detection limit for stool samples was 10⁵ gc/mL, thus bettering detection limits of commercially available norovirus diagnosis quick kits of 100-fold; the detection limit in spiked mussels however was ten-fold higher than in stool samples. Further assays showed a decrease in fluorescence intensity for heat- or UV-inactivated virus particles. Overall, we demonstrate the application of a flow cytometry approach for direct detection of small non-enveloped virus particles such as noroviruses. An adaptation of the technology to routine diagnostics has the potential to contribute a rapid and sensitive tool to norovirus outbreak investigations. Further improvements to the method, notably decreasing the detection limit of the approach, may allow the analysis of naturally contaminated food and environmental samples.

Graphic Abstract

中文翻译：

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开发一种特异性抗衣壳抗体和基于磁珠的免疫测定法，通过流式细胞术检测粪便样品和尖刺贻贝中的人类诺如病毒颗粒

人类诺如病毒在全球范围内造成了相当大的健康负担。在这里，使用抗体包被的磁珠开发了一种流式细胞仪方法，用于检测生物废物和食品样品中的它们。产生针对鼠诺如病毒和各种人类诺如病毒基因型的抗肽抗体用于捕获并包被在磁珠上。然后实施流式细胞术测定以检测患者粪便样本和加标诺如病毒的贻贝消化组织中的珠结合人诺如病毒 GI.3。粪便样本的检出限为 10 ⁵ gc/mL，从而将市售诺如病毒诊断快速试剂盒的检测限提高100倍；然而，加标贻贝的检测限是粪便样本的 10 倍。进一步的测定显示热灭活或紫外线灭活的病毒颗粒的荧光强度降低。总体而言，我们展示了流式细胞术方法在直接检测小的无包膜病毒颗粒（如诺如病毒）中的应用。将该技术应用于常规诊断有可能为诺如病毒爆发调查提供快速和敏感的工具。对该方法的进一步改进，特别是降低该方法的检测限，可能允许分析自然污染的食品和环境样品。

图形摘要