Department of Biological Systems Engineering, Virginia Tech, Blacksburg, Virginia 24061, United States.
Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Tech, Blacksburg, Virginia 24061, United States.
J Agric Food Chem. 2023 Jun 7;71(22):8665-8672. doi: 10.1021/acs.jafc.3c01946. Epub 2023 May 25.
Human noroviruses pose grave threats to public health and economy. In this study, we genetically engineered yeast ( EBY100) to display specific norovirus-binding nanobodies (Nano-26 and Nano-85) on cell surface to facilitate the concentration of noroviruses for improved detection. Binding of norovirus virus-like particles (VLPs) to these nanobody-displaying yeasts was confirmed and characterized using confocal microscopy and flow cytometry. The ability of our engineered yeasts to capture norovirus VLPs can reach up to 91.3%. Furthermore, this approach was applied to concentrate and detect norovirus VLPs in a real food matrix. A wide linear detection range (1-10 pg/g) was observed, and the detection limit on spiked spinach was calculated as low as 0.071 pg/g. Overall, our engineered yeasts could be a promising approach to concentrate and purify noroviruses in food samples for easy detection, which allows us to prevent the spread of food-borne virus in the food supply chain.
人类诺如病毒对公共卫生和经济构成严重威胁。在本研究中，我们通过基因工程改造酵母（EBY100），使其在细胞表面展示特定的诺如病毒结合纳米抗体（Nano-26 和 Nano-85），以便于浓缩诺如病毒，从而提高检测效率。使用共聚焦显微镜和流式细胞术证实并表征了这些纳米抗体展示酵母与诺如病毒病毒样颗粒（VLPs）的结合。我们工程化酵母捕获诺如病毒 VLPs 的能力最高可达 91.3%。此外，该方法还应用于浓缩和检测真实食品基质中的诺如病毒 VLPs。观察到宽线性检测范围（1-10 pg/g），并且在添加菠菜上的检测限计算低至 0.071 pg/g。总体而言，我们的工程化酵母可以作为一种有前途的方法，用于浓缩和纯化食品样本中的诺如病毒，以便于检测，从而防止食源性病毒在食品供应链中的传播。
