Development Of Immunochromatographic Analysis For Acidovorax Avenae Subsp. Citrulli

The Acidovorax avenae subsp.citrulli(Aac)and P.syringae pv.lachryrias are two important pathogens for cucumis melo,they have very important impact on the global melon industry.Because the two kinds of diseases have the very similar symptoms,it is very difficult to distinct them or prevent them in the fields.In order to efficiently and accurately distinguish the two diseases in the field,this topic using the monoclonal antibody and polyclonal antibody of Aac,which based on the principle of double antibodies sandwich,built a kind of immune chromatography test paper and established a highly efficient gold immune chromatography analysis method.By this method,we make the detection of Aac become rapidly and accurately in the fields.The immune chromatography test paper based on the principle of specific binding between antigen and antibody,has the more rapidly and efficient advantages compared with the traditional molecular detection methods.In this paper,we will introduce the development and testing effect of the immune chromatography test paper from the flowing aspects:(1)We screened two hybridoma cell lines of producting monoclonal antibody(McAb)stably against Acidovorax avenae subsp.citrulli(Aac).Besides,we obtained antiserum of polyclonal antibody(PcAb).And we obtained the McAb and PcAb by purified which were the materials of establishing the immunoassay.(2)We successfully built a immunochromatography assay(ICA)method to detect the Acidovorax avenae subsp.citrulli(Aac)in the field.The immunochromatography assay(ICA)method based on double antibody sandwich theory,in which the colloidal gold-antibody 5A3 as identify antibody,the polyclonal antibodies 9154 as capture antibodies.And we optimized the parameters and working solution of the ICA test strip.In the qualitative detection,the minimum concentration of Aac was 1×105cfu/ml,we can easier find it on the ICA test strip by naked.Moreover,the determination of Aac has strong specificity and without cross-reaction with P.syringae pv.lachrymas.From above we can see that the developed ICA test strip can be used to rapidly and specifically detect the Aac in field,which is helpful for the development of melon and other melon planting industry.(3)We built a new theoretical method for quickly and efficient Acidovorcax avenae subsp.citrulli(Aac)detection,based on surface-enhanced Raman scattering theory.