间接ELISA检测鸡大肠埃希氏菌菌毛抗体方法的建立

Development of an indirect ELISA for detecting antibody to pili of Escherichia coli of ckick origin

用大肠埃希氏菌O1、O2 、O3、O78和O89株制备ELISA抗原 ,与抗鸡Ig单抗 1B7酶结合物建立了检测鸡血清抗体水平的间接ELISA方法。经交叉试验、阻断试验、重复性试验等表明该方法重复性好、特异性强、灵敏度高 ;确立了将鸡血清 10 0倍稀释检测ELISA效价 (ET)的回归方法 y =2 .45 3 + 0 .131x(P >0 .0 5 ) ,可用于定量测定。用IH与ELISA方法的比较试验表明 ,ELISA法比IH法敏感性高 4— 10

An indirect ELISA method for measuring antibody titer to pili of E.coli of chick origin was established with E.coli strains O 1、O 2、O 78 and O 89 prepared antigen for ELISA and anti prepared antigen for ELISA and anti chick Ig monoclonal antibody 1B7 enzyme conjugate, which was confirmed to have better repeat, specificity and higher sensitivity by cross tests, block tests and reproductive tests. The regressive equation that calculates an ELISA titer in 100 times dilution chicks sera is affirmed as Y=2.453+131X which is able to quantify. The comparisons between indirect hemagglutination (IH) test and the ELISA revealed that the ELISA test was 4 to 100 times more sensitive than IH.