Development of A Monoclonal Antibody-Coated Immunomagnetic Beads for Separation and Detection of Norovirus (Genogroup II) in Oysters.

The present study aimed to develop and evaluate a reverse transcription- polymerase chain reaction (RT-PCR) assay that incorporated immunomagnetic beads (IMS/RT-PCR) coated with a monoclonal antibody (Mab) for the separation and detection of noroviruses (NVs, genogroup II) in oysters. Mab-coated beads and polyclonal antibody (Pab)-coated beads were added to oyster extracts containing a series of tenfold dilution of NVs-positive fecal extracts. The oyster extracts were also treated with Proteinase K method. RNA was extracted and used as template for RT-PCR. Mab-based IMS/RT-PCR could detecte 10 RT PCR unit in oyster homogenate, it was 10 times more sensitive than Pab-based IMSIRT-PCR, and 10² times more sensitive than proteinase KIRT-PCR. Analysis of 473 oysters showed that Mab-based IMS/RTPCR has the highest percentage of positive samples (5.29%), followed by Pab-based IMS/RT-PCR (3.81%), and proteinase KIRT-PCR (2.11%). Mab-based IMS/RT-PCR is a sensitive method for the detection of NVs in oysters and could be a good candidate for routine screening and risk assessment. [ABSTRACT FROM AUTHOR]