›› 2015, Vol. 42 ›› Issue (6): 1383-1388.doi: 10.16431/j.cnki.1671-7236.2015.06.009

Previous Articles     Next Articles

Establishment of Indirect ELISA Method for Detecting Antibodies against Avain Infectious Bronchitis Virus

ZHENG Wei-feng, YAN Fang, WANG Pan, ZHAO Yu-jun, GAO Wen-wei, MA Hai-li, LI Xu-ying, NING Guan-bao, TIAN Wen-xia, LIANG Hui-xia, LIU Lin   

  1. Veterinary Microbiology and Immunology Laboratory, College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China
  • Received:2014-11-10 Online:2015-06-20 Published:2015-07-23

Abstract: The purpose of this research was to establish a rapid detection serological method against avain infectious bronchitis virus (IBV).In this study,an indirect ELISA method was established using IBV as the detected antigen and a variety of testing conditions were optimized.The results showed that the optimal antigen coating concentration was 19.2 μg/mL and the optimal condition for coating was incubated at 37 ℃ for 1 h and then 4 ℃ overnight.The optimal dilutions of serum and enzyme labeled antibody were 1:800 and 1:7 000 incubated at 37 ℃ for 60 min,respectively.The optimal condition of chromogenic substrate was incubated at 37 ℃ for 10 min in the dark.The specificity,repeatability and sensitivity tests proved that the indirect ELISA did not cross-react with positive antiviral sera of other chicken diseases,had good repeatability and could detect IBV antibody when serum was diluted 1:12 800.We concluded that the established indirect ELISA was specific,repeatable and sensitive.

Key words: avian infectious bronchitis virus (IBV); indirect ELISA; antibody detection

CLC Number: