F18+ETEC分离株的鉴定·菌毛蛋白提取及生物活性验证

目的]探讨F18+ETEC分离株的鉴定、菌毛蛋白的提取及菌毛蛋白生物活性的初步验证。方法]分别以分离株A20、D20为模板进行PCR扩增,用1%琼脂糖凝胶电泳检测PCR产物。采用热抽提法分别提取F18标准株和分离株A20、D20的菌毛蛋白,并分别制备弗氏佐剂苗,免疫产蛋鸡,采用间接ELISA法定期检测母鸡的IgY和血清抗体效价。结果]采用TSB培养基,37℃摇床培养24 h,分离株和标准株菌毛均获得了良好的表达。分离株A20、D20经PCR扩增、琼脂糖凝胶电泳、SDS-PAGE方法鉴定均为F18+ETEC。F18ab分离株和标准株的菌毛对产蛋鸡均具有良好的免疫原性,卵黄中IgY效价最高可达1∶25 600,血清中抗体效价高于卵黄中IgY效价,最高可达1∶51 200。结论]分离株A20、D20均为F18+ETEC,且对产蛋鸡均具有良好的免疫原性。

Identification of F18+ ETEC Isolates,Extraction of Their Fimbriae Proteins and Preliminary Verification of Their Biological Activity

Objective] The purpose was to discuss the identification of F18+ ETEC isolates,extraction of their imbriae proteins from these strains and preliminary identification of their biological activities.Method] With isolates A20 and D20 as the templates,the PCR amplification was carried out and the PCR products were detected by 1% agarose gel electrophoresis.The fimbriae proteins were extracted from standard strains F18 and isolates A20 and D20 by thermal extraction method to prepare Freund's adjuvant vaccine a...