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通过原子力显微镜和病毒回收测定MS2与食品接触表面的差异相互作用

Differential MS2 Interaction with Food Contact Surfaces Determined by Atomic Force Microscopy and Virus Recovery.

作者信息

Shim J, Stewart D S, Nikolov A D, Wasan D T, Wang R, Yan R, Shieh Y C

机构信息

Illinois Institute of Technology, Department of Chemical and Biological Engineering, Chicago, Illinois, USA.

U.S. Food and Drug Administration, Bedford Park, Illinois, USA.

出版信息

Appl Environ Microbiol. 2017 Dec 1;83(24). doi: 10.1128/AEM.01881-17. Print 2017 Dec 15.

Abstract

Enteric viruses are recognized as major etiologies of U.S. foodborne infections. These viruses are easily transmitted via food contact surfaces. Understanding virus interactions with surfaces may facilitate the development of improved means for their removal, thus reducing transmission. Using MS2 coliphage as a virus surrogate, the strength of virus adhesion to common food processing and preparation surfaces of polyvinyl chloride (PVC) and glass was assessed by atomic force microscopy (AFM) and virus recovery assays. The interaction forces of MS2 with various surfaces were measured from adhesion peaks in force-distance curves registered using a spherical bead probe preconjugated with MS2 particles. MS2 in phosphate-buffered saline (PBS) demonstrated approximately 5 times less adhesion force to glass (0.54 nN) than to PVC (2.87 nN) ( < 0.0001). This was consistent with the virus recovery data, which showed 1.4-fold fewer virus PFU recovered from PVC than from glass after identical inoculations and 24 h of cold storage. The difference in adhesion was ascribed to both intrinsic chemical characteristics and the substrate surface porosity (smooth glass versus porous PVC). Incorporating a surfactant micellar solution of sodium dodecyl sulfate (SDS) into the PBS reduced the adhesion force for PVC (∼0 nN) and consistently increased virus recovery by 19%. With direct and indirect evidence of virus adhesion, this study illustrated a two-way assessment of virus adhesion for the initial evaluation of potential means to mitigate virus adhesion to food contact surfaces. The spread of foodborne viruses is likely associated with their adhesive nature. Virus attachment on food contact surfaces has been evaluated by quantitating virus recoveries from inoculated surfaces. This study aimed to evaluate the microenvironment in which nanometer-sized viruses interact with food contact surfaces and to compare the virus adhesion differences using AFM. The virus surrogate MS2 demonstrated less adhesion force to glass than to PVC via AFM, with the force-contributing factors including the intrinsic nature and the topography of the contact surfaces. This adhesion finding is consistent with the virus recoveries, which were determined indirectly. Greater numbers of viruses were recovered from glass than from PVC, after application at the same levels. The stronger MS2 adhesion onto PVC could be interrupted by incorporating a surfactant during the interaction between the virus and the contact surface. This study increases our understanding of the virus adhesion microenvironment and indicates ways to mitigate virus adhesion onto contact surfaces.

摘要

肠道病毒被认为是美国食源性感染的主要病因。这些病毒很容易通过食品接触表面传播。了解病毒与表面的相互作用可能有助于开发更好的去除方法,从而减少传播。以MS2噬菌体作为病毒替代物,通过原子力显微镜(AFM)和病毒回收试验评估了病毒对聚氯乙烯(PVC)和玻璃等常见食品加工和制备表面的粘附强度。使用预结合有MS2颗粒的球形珠探针,从力-距离曲线中的粘附峰测量MS2与各种表面的相互作用力。磷酸盐缓冲盐水(PBS)中的MS2对玻璃(0.54 nN)的粘附力比对PVC(2.87 nN)的粘附力低约5倍(<0.0001)。这与病毒回收数据一致,该数据显示,在相同接种和冷藏24小时后,从PVC中回收的病毒空斑形成单位(PFU)比从玻璃中回收的少1.4倍。粘附力的差异归因于内在化学特性和底物表面孔隙率(光滑的玻璃与多孔的PVC)。将十二烷基硫酸钠(SDS)表面活性剂胶束溶液加入PBS中可降低PVC的粘附力(~0 nN),并使病毒回收率持续提高19%。通过病毒粘附的直接和间接证据,本研究说明了对病毒粘附的双向评估,用于初步评估减轻病毒对食品接触表面粘附的潜在方法。食源性病毒的传播可能与其粘附性质有关。通过定量接种表面的病毒回收率来评估病毒在食品接触表面的附着情况。本研究旨在评估纳米级病毒与食品接触表面相互作用的微环境,并使用AFM比较病毒粘附差异。通过AFM,病毒替代物MS2对玻璃的粘附力比对PVC的粘附力小,其力贡献因素包括接触表面的内在性质和形貌。这一粘附发现与间接测定的病毒回收率一致。在相同施加水平后,从玻璃中回收的病毒数量比从PVC中回收的病毒数量更多。在病毒与接触表面相互作用期间加入表面活性剂可中断MS2对PVC的更强粘附。本研究增进了我们对病毒粘附微环境的理解,并指出了减轻病毒对接触表面粘附的方法。

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