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

摘要

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 IMS/RT-PCR, and 10~2 times more sensitive than proteinase K/RT-PCR. Analysis of 473 oysters showed that Mab-based IMS/RT-PCR has the highest percentage of positive samples (5.29%), followed by Pab-based IMS/RT-PCR (3.81%), and proteinase K/RT-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.