RESUMO
This study was aimed to develop a sandwich ELISA kit for the diagnosis of bovine tuberculosis.And it was applied and evaluated in the quarantine of bovine tuberculosis.We established a bovine IFN-γ release method in vitro and developing three batches of kits.The sensitivity,repeatability and retention period of the kit were all evaluated.Totally 961 serum samples were tested using the developed sandwich ELISA kit tuberculin skin test and a commercial ELISA kit.Our results showed that the detection limit of this ELISA was 8.21 mg/mL.The repeatability tests showed good reproducibility in the intraassay and inter-assay.At the same time,the retention period of the kit was more than 12 months.Compared with the tuberculin skin test,the positive coincidence rate was 70.59％ and the negative coincidence rate was 99.20％,while the total coincidence rate was 98.44％.And compared with the BOVIGAMTM kit,the positive coincidence rate was 91.30％ and the negative coincidence rate was 99.78％,while the total coincidence rate reached 99.58％.At the same time,the sensitivity and specificity of the sandwich kit were 85.00％ and 100％,respectively.We established a bovine IFN-γ release method in vitro and developing corresponding kits successfully have a good application prospect.