Evaluation of Porcine Gastric Mucin Magnetic Bead Enrichment for the Detection of Norovirus in Bivalve Mollusks
Journal Title: International Journal of Research in Agricultural Sciences - Year 2018, Vol 5, Issue 2
Abstract
Norovirus (NoV) represents one of the leading causes of foodborne illness worldwide. Standard methods for detection or quantification of NoV in bivalve mollusks are now available, but cannot discriminate between infectious and non-infectious virus particles. Based on the ability of intact NoV particle to bind to porcine gastric mucins conjugated to magnetic beads (PGM-MB), this study aimed to develop an enrichment method allowing to assess the infectivity of NoV particles in bivalve mollusks. To achieve this aim, we have investigated the effects of pH of binding buffer, amounts of PGM-MB and the use of additives on the recovery of NoV. The optimal parameters for the capture of NoV in modified Britton–Robinson buffer were pH 6.0; presence of 5% glycerol and 100 µl of PGM-MB. To avoid the negative effect of oyster matrix on the capture of NoV, oyster homogenate needs to be diluted three times in binding buffer. This optimal procedure allowed the detection of 2 × 102 NoV genome copies/ml in clean system and 5 × 103 NoV genome copies/g of oyster digestive tissues. The advantages of this new method are its simplicity and capability of assessing the infectivity of NoV particles.
Authors and Affiliations
Hang Thu Ngo, et al.
Keywords
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