Development And Application Of The Solid-phase Competition ELISA And Double Antiboby Sandwich ELISA For Detection Of Serotype O FMD

Foot-and-mouth Disease(FMD) is a highly contagious and devastating viral disease of cloven-hoofed animals, it Seriously affect the development of animal husbandry in the world. Prevention and control of FMD in developed countries is mainly to kill infected animals, our country mainly vaccine immunization. The rapid diagnosis of FMD is very important for the prevention and control of FMD.In order to make a rapid and specific diagnosis of FMD, we obtain mouse monoclonal antibody for the typing of FMDV serotype O and prepare a large number of ascites, and the antibody was purified and labeled with HRP, which was named as 2B5-HRP. A Solid-phase competition ELSA(SPCE) was developed using 2B5-HRP. The results showed that we succeed obtain antibody for the typing of FMDV serotype O and determine examining test at 1:8 dilution with a cut-off point of 40 percentage inhibition(PI) of reaction. Sensitivity and specificity is an important indicator for the evaluation of diagnostic methods, The specificity of SPCE was 97.8% and the sensitivity of SPCE was 98.2% using 176 positive sera and 273 negative sera, respectively. The coincidence rate of SPCE, LPB-ELISA, VDPro and PrioCHECK were higher than 95%. The coefficient of variability of within-run and between-runs were well, and the levels of SPCE and LPB-ELISA in the detection of immune animal antibody were basically consistent. The sensitivity and specificity of the SPCE is better, it can specificly diagnose typing of FMDV serotype O and can be used for the large-scale detection of FMD, provides a new means for the epidemiological investigation of FMD.To establish rapid detection for foot-and-mouth virus(FMDV), a double antibody sandwich ELISA for FMDV was developed using rabbit monoclonal antibody and 2B5-HRP,respectively. The results showed that the sandwich ELISA was specific for O type FMDV,but no cross reaction with swine vesicular disease virus, FMDV type of A and Asia-I. The sensitivity and specificity between sandwich ELISA and RT-PCR were basically consistent. The coefficient of variability of within-run and between-run were better. The andwich ELISA detection method established in this study can be used for the detection of FMDV type of O, which provides a technical means for the epidemiological investigation and control of FMDV.