致仔猪水肿病大肠杆菌快速检测方法的建立

Establishment of a Diagnostic Method for Rapid Detection of Edema Disease in Piglets Caused by E.coli

将水肿病菌株S在TSB培养基中连续传代两次 ,离心后用生理盐水洗涤 ,制成1×108的细菌悬液。菌液倍比稀释 ,分别与适当浓度的F107菌毛单抗作玻板凝集、间接ELISA、斑点 -ELISA试验 ,结果表明 ,当单抗100×稀释时 ,三种方法的细菌最小检出量分别为2.5×107、6×105、1×103,斑点 -ELISA最敏感。从患水肿病的仔猪小肠黏膜上刮取黏液 ,溶于PBS中 ,将此溶液一部分稀释后在麦康凯平板上菌落计数 ,另一部分作为抗原做斑点 -ELISA。结果显示致病菌在小肠黏膜上大量存在 ,斑点 -ELISA能敏感、快速的检测出来。这一检测方法的建立 ,为其它大肠杆菌病的诊断提供了较好的借鉴。

One strain S of E.coli from weaned pigs with edema which possess fimbriae F107was cultured in trypticase soy broth(TSB)for24hours,at37℃.The strain reacted with anti-fimbriae F107Mab by slide agglutinaˉtion,indirect-ELISA and Dot-ELISA.Results showed that the strain could be identified by Dot-ELISA sensiˉtively.According to adhesive character of fimbriae F107,we isolated strains from the mucosal of the small intesine and identified these bacterium with anti-fimbriae F107antibodies by Dot-ELISA.The results showed that Dot-ELISA could detect the strains that expressed fimbriae sensitively.Major advantage of the technique was sensitive and prompt.The Dot-ELISA with Mab had potential uses as a powerful diagnostic technique for ED.