| New Castle disease is an acute and highly contagious disease caused by Newcastle disease virus, which causes high rate of infection and death and great economic losses to the poultry industry, especially to chicken industry. Tri-sodium citrate with aqueous gold chloride are warmed up and mixed to make gold sol. Colloidal gold particles are coupled with antibody or antigen using to diagnose antigen or antibody. On one hand, Monoclonal antibody against NDV is prepared to manufacture immunochromatography test strip on diagnosing antigen, and the strip has strong specificity. On the other hand, a semi-quantitative run test strip for antibody is manufactured, and the strip has high sensitivity.Firstly, NDV separation and purification are used by the method of different velocity and chromatography SephacrylTM S-200 for next step. And its purity is very good, which is prepared for the antigen of monoclonal antibody and polyclonal antibody. With the method of n-Caprylic acid-ammonium sulphate the serum antibody of rabbits gainst ND is taken. Purity of IgG is identified by SDS-PAGE.The key of the experiment is the preparation of colloidal gold. By repeat test, the optimization as follows: diameter of colloidal gold particle is 20nm; the combination quantity of colloidal gold with antigen and antibody is 36μg/ml,the pH is 9 and 8.2. At the same time, the check line and the contrast line are sprinkled 0.6mg/ml and 0.5mg/ml protein when makes the strip.
|
PDF Full Text Request