Laboratory of Food Microbiology and Food Preservation, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, B-9000 Ghent, Belgium.
J Virol Methods. 2011 Nov;177(2):153-9. doi: 10.1016/j.jviromet.2011.07.013. Epub 2011 Aug 9.
Attempts were made to discriminate between infectious and non-infectious Noroviruses (NoVs) based on their viral binding properties followed by reverse transcription polymerase chain reaction (RT-PCR). Murine norovirus-1 (MNV-1) was employed as a surrogate to test the principle. Detection of both infectious and inactivated MNV-1 was investigated by the plaque assay, RT-PCR and binding-based RT-PCRs. The cell line RAW 264.7 and the ganglioside GD1a were used as binding receptors respectively in combination with RT-PCR. In the second stage of testing, similar approaches were applied to the two main genogroups of human NoVs (GI and GII). Differentiated Caco-2 cells and pig gastric mucin were tested as the binding receptors. Bovine serum albumin (BSA) was used as a non-specific binding control. In this study, the binding-based RT-PCRs decreased the detection of non-infectious NoVs by 1-3-log(10) while all infectious viral particles were detected. No significant difference was observed between the binding-based RT-PCRs within the concentration range investigated, except the binding level of human NoVs GII to pig gastric mucin was higher than to differentiated Caco-2 cells and BSA. This study indicates an improvement in the evaluation of the infectivity of non-cultivable human NoVs. This is also a comprehensive study on both specific and non-specific binding properties of NoVs.
尝试根据病毒结合特性对传染性和非传染性诺如病毒（NoV）进行区分，然后进行逆转录聚合酶链反应（RT-PCR）。采用鼠诺如病毒-1（MNV-1）作为替代物来测试该原理。通过噬斑测定、RT-PCR 和基于结合的 RT-PCR 检测到传染性和失活的 MNV-1。RAW 264.7 细胞系和神经节苷脂 GD1a 分别用作结合受体，并与 RT-PCR 结合使用。在测试的第二阶段，类似的方法应用于人类诺如病毒的两个主要基因型（GI 和 GII）。分化的 Caco-2 细胞和猪胃粘蛋白被测试为结合受体。牛血清白蛋白（BSA）用作非特异性结合对照。在这项研究中，基于结合的 RT-PCR 将非传染性 NoV 的检测降低了 1-3 个对数（10），而所有传染性病毒颗粒均被检测到。除了人 NoV GII 对猪胃粘蛋白的结合水平高于分化的 Caco-2 细胞和 BSA 外，在所研究的浓度范围内，基于结合的 RT-PCR 之间未观察到显著差异。本研究表明，对不可培养的人 NoV 的感染性评估有所提高。这也是对 NoV 的特异性和非特异性结合特性的综合研究。
