Study On The Muscovy Duck Reovirus Disease's Diagnosis Technology

Indirect Enzyme-Linked Immunosorbent Assay(ELISA) and Latex Agglutination Test(LAT) were studied and applied to diagnose the muscovy duck reovirus disease. The muscovy duck reovirus , which was separated and appraised by professor Wu,was use in the two test.1. The virus was cultivated in the cell of muscovy duck embryo. The antigen was prepared for ELISA and LAT by centrifugation and precipitation. TCIDso of the virus was detected and calculated. The result was that per millimeter cell fluid consisted of 56990 pieces of TCIDso. The virus protein content was 5.628mg/ml.2. The first polyclone-antibody of the virus and the second polyclone-antibody were prepared. The healthy muscovy ducks were immunized four times(one time per ten days). The result indicated that the positive price was as much as 1:16. The positive serum was purified by precipitation and ion -exchange chromatography. The IgG protein content was detected as much as 4.242mg/ml. On the other, the anti-muscovy duck poly-antibody of rabbit was prepared, which positive price was 1:8 and the protein content was 5.395mg/ml. Then the HRP was marked to the purity rabbit anti-muscovy duck IgG. The marked rate was 0.6909.3. The reaction condition of indirect ELISA was succeeded in developing through a lot of experiment. The results indicated that diluting the antigen to 0.0704mg/ml and the serum to 1:40 and the marked IgG to 1:200 were the best reaction condition. The antigen's specificity , sensitivity and stability were tested. The result meaned that the Indirect ELISA had good speciality, sensivity and stability. It is suitable for serological epidemiological survey and serological diagnosis of muscovy duck reovirus disease.4. The reaction condition of Latex Agglutination Test (LAT) was soluted through a lot of experiment. The best suitable condition for coating the latex was determined by titration test. The result was that antibody content 0.4242 mg/ml, reaction temperature 37 C and reaction time 2 hours were the best reaction conditions o The LAT had good speciality, sensivity and stability. The LAT detected the Muscovy Duck Reovirus from the stool of the infection duck on the third day after the artificial infection. And it also detected the virus from the spleen and the liver on the fourth and the fifth day after infection. But it was negative from the heart of the dead duck.