Development Of An Indirect ELISA For YAN BIAN Cattle's Clostridium Welchii Disease Of D

Clostridium welchii is one of condition pathogenic bacterium with allthe word. In condion, it can produce disease. It is morbidity area in YANBIAN. YAN BIAN Cattle's Clostridium welchii's disease cause toClostridium welchii of D. Thallus of Clostridiumwelchii has whole thallusantige.The various factors and conditions for indirect ELISA were explored,and the optimal reaction conditions were determined. It was shown thatthe optimal concentration of Clostridium welchii's thallus antige forcoating of plate was 50μg/ml, the optimal coating condition of recombinantprotein was 37℃1h, the dilution of serum sample was 1:100, the workingconcentration of HRP-labeled rabiit anti-bovine IgG was 1:1000, serumsample for detecting and rabbit anti-bovine IgG should be incubated at37℃for 1h before terminated with the stopping solution. The optimalconfining solution and confining time were 3ï¼…skimmed milk of PBST, 37℃for lh respectively. The substrate for indirect ELISA was incubated atRT for 25 min before reading the OD₄₀₅ The threshold for indirect ELISAwas 0.032. In addition, 100 bovine serum samples collected from Yanbiancanton were detected by the indirect ELISA and found the positive rateis 11ï¼….In the domestic field of Clostridium welchii's thallusantige, thistest firstly successfully apply Clostridium welchii's thallus antige asan antigen todeveloped indirect ELISA to detect Clostridium welchii'sdisease. It should afford a rapid, sensitive and specific means of serodiagnosis in monitoring Clostridium welchii's disease infection orepidemiology investigation in the field.