| :The different diameter colloidal gold particles were synthesized by reducing hydrogen tetrachloroaurate with sodium citrate using the electric cooker which was a heating up installation.At the same time, we gained the referenced data which prepared colloidal gold particles form 10 to 50nm. The 20nm colloidal gold particles were selected,according to the different diameter colloidal gold particles showing different color(easy to judge). The TMV was purified on the zonal ultracentrifuging at 4℃,then use the method of colloidal gold immunoelectronmicroscopy to detect TMV. The result showed that the TMV had been purified. Using the TMV immunized rabbit, the received serum was purified by caprilic acid/ammonium sulfa mate precipitation and DEAE Sephadex A50 hydronium exchanging.The purified antibodies were analyzed by the conventional PAGE and agar diffusion test.And the result showed that the IgG in high purity and good activities.The gold sol was adjusted to pH 8.0 then at 4℃, the labeled rabbit anti-TMV IgG was purify on zonal ultracentrifuging. We diluted the labeled rabbit anti-TMV IgG in PBS (containing I%BSA) at the rate of 1:2. The labeled antibodies was immobilized to nitrocellulose membrane(NC). The two kind of antibody concentration was optimized by the square matrix test. Meanwhile, a optimized and stable formula was obtained. The test strip was placed in 4℃desiccator,after 5 month its effect of detection and releasing still hold the line.40 clinical samples were detected by the strip comparing with Agdia Immunostrip Test.Our products’ results accord to Agdia Immunostrip Test.The test strip show is easy to be justified;time-saving,sensitive,and suitable for at grassroot level and epidemiology investigate.These key techniques and the date will help us research and develop a series of GICA strip to detect the most floristic virus. |
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