Simultaneous Detection and Genotyping of Norwalk-like virus (GⅡ) by blot hybridization

摘要

Seven Norwalk-like viruses (NLVs): 2 from stool samples and 5 from 52 oyster samples during 2002-2004 have been detected by using RT-PCR to amplify the segment of the RNA dependent polymerase-coding gene. A probe was designed based on sequence analysis and has been labeled by biotin for confirmation and genotyping of the samples by blot hybridization. The blot hybridization assay, which provides a simple and quick method to confirm the result of PCR, shows that the biotin labeled probe could detect the plasmids of the 7 detected samples, but not with other species. The hybridization with PCR products also showed the same result of plasmids. Comparing the sensitivity between RT-PCR and blot hybridization. These data demonstrated that the established blot hybridization was a quick method and is sensitive and specific for detection of Norwalk-like viruses and could simultaneously genotype the detected samples.