Establishment Of A Double Antibody Sandwich ELISA And Colloidal Gold Immunochromatographic Strip For The Detection Of Infectious Bronchitis Virus

A double antibodies sandwich ELISA was established using monoclonal antibody 2D6 as capture antibody and 11C10 labeled with horseradish peroxidase as enzyme-labeled antibody to test IBV. All of the reaction conditions were optimized. The DAS-ELISA only test IBV had positive result and had no cross-reaction with other chicken infection virus. The allantoic fluid of IBV was diluted 160 times then the DAS-ELISA still able to detect it.Colloidal gold of 25nm was prepared, by trisodium citrate reduction method. The purified monoclonal antibody 11C10 labeled with colloidal gold was adsorbed on the glass fiber membrane as gold-labeled pad. The purified monoclonal antibody 2D6 and a sleep anti-mouse IgG antibody was respectively coated on the test and control line of the nitrocellulose membrane. Then the colloidal gold immunochromatographic strip for the rapid detection of IBV was assembled by gold-labeled pad, nitrocellulose membrance and absorbent paper. Each step were optimized during the developed of the strip. The allantoic fluid of IBV was diluted 80 times then the strip still able to detect it and had no cross-reaction with other chicken infection virus.The strip and DAS-ELISA detected the virus in the 55 clinical samples taken from chicken with suspected IBV infection. The results coincidence rate was 92.73%, indicating that the strip was a suitable method for the diagnosising and detecting of IBV.