Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 177, Issue 2, Pages 153-159Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2011.07.013
Keywords
Norovirus; MNV-1; RT-PCR; Binding; Infectivity; Inactivation
Funding
- Ghent University
- China Scholarship Council
- Research Foundation-Flanders (Fonds voor Wetenschappelijk Onderzoek [FWO] Vlaanderen)
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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 GDla 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 (Cl 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. (C) 2011 Elsevier B.V. All rights reserved.
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