Detection Of Norovirus In Water Based On Nanomaterial

Norovirus is an important foodborne and water-borne virus,which often causes an outbreak of acute gastroenteritis by contaminating water sources.The identification of Norovirus is mainly through the real-time RT-PCR method,because Norovirus can not be cultured in the laboratory by cell and tissue culture,and can not be detected when the amount of Norovirus in the water is low.In order to overcome the problem of enrichment of noroviruses in water,this paper proposes to establish a method for enrichment of hydroviruses with nanometal oxides(titania,zirconia)modified cellulose membranes,and optimize the parameters for specific construction and detection.Finally,a method of fast,efficient,simple and cheap enrichment detection is established to solve the problem of Norovirus detection technology in water.(1)A real-time fluorescence quantitative RT-PCR method was established to detect Norovirus.The sensitivity,specificity and stability of the method were verified.The actual detection effect of the method was tested by clinical samples.The standard curve was drawn with GII Norovirus standard plasmid.The regression equation was Y=-3.3661 x +46.441 and the correlation coefficient R2 was 0.9993.The lowest detection concentration was 9.1×10~2 copies/mu L.The variation coefficients of CT values in three concentrations were less than 2%.The method had high sensitivity,accuracy and reproducibility.It can be applied to the detection of water,such as virus enrichment.(2)The metal oxide nanomaterials were modified to the surface of cellulose membrane by sol-gel method,and the enrichment effects of the two materials were tested.Comparing the results,it is found that the nano titanium dioxide modified filter paper has better enrichment effect and can be used in the detection method of norovirus in water.(3)The conditions for enriching the Norovirus in water with nano-titanium dioxide modified membrane were optimized.When the concentration of Norovirus in the water is9.1×103copies/?L,enrich with 3L nano-Ti O2 modified filter paper with 10 L water sample,the flow rate is controlled at 0.5L/min,and it contains(3% beef extract and 0.1M glycine,The best effect on the elution of the p H 9.5 eluate is to increase the virus concentration in the water to 3.9×10~6 copies/?L.Concentrated by 429 times,the method recovery was 4.3%.(4)By comparing the anion membrane method and the flocculation precipitation method,the nano-titanium dioxide modified filter paper enrichment method has the characteristics of large concentration and large volume.Under certain conditions,more Norovirus can be enriched for real-time fluorescence quantitative RT-PCR detection..The detection of 54 actual samples and the detection of 4 GII Noroviruses showed that the method has certain practical value.