Detection of waterborne norovirus with realtime fluorescense quantitative RT-PCR

Objective To establish a method for waterborn norovirus enrichment and detection with realtime reverse transcription(RT)-PCR.Methods Norovirus positive fecal samples were diluted gradiently with purified water;the simulated water samples were filtered through a microbial purification system with cation mixed cellulose acetate membrane and added with 20%PEG6000;then the virions in the samples were obtained with two times of high-speed centrifugation and the nucleic acids of the virus were extracted and detected with fluorescence quantitative realtime RT-PCR.Results Fluorescence quantitative realtime RT-PCR revealed a good amplification of recombinant plasmid (T-NVS)with a linear correlation of 98%;the detection limit of the established method for norovirus was 100 copies/μL.After the enrichment,the virus recovery rate ranged form 5.75% to 19.74% for the simulated water samples with the dilution ratios of 10 to 1 000.Conclusion A method for enrichment and detection of norovirus in water samples was preliminarily established.