牛结核病鉴别诊断的方法研究

被引：12

作者：

余大海

韦一然

蔡宏

朱玉贤

机构：

[1] 北京大学蛋白质工程及植物基因工程国家重点实验室

来源：

中国人兽共患病学报 | 2006年 / 10期

关键词：

牛结核病; TB10.4蛋白; 间接ELISA法;

D O I：

暂无

中图分类号：

S854.44 [];

学科分类号：

0906 ;

摘要：

目的构建含结核分枝杆菌(Mycobacterium tuberculosis)tb10.4基因片断的原核表达载体,在E.coli中诱导表达带组氨酸标记的该融合蛋白。分别以TB10.4蛋白,PPD蛋白以及TB10.4与MPT83混合蛋白为抗原,用间接ELISA法鉴别牛结核病。方法PCR法扩增tb10.4基因片段,连接到pET-22b(+)原核表达载体中,将重组子转化到大肠杆菌蛋白酶缺陷型菌株BL21(DE3)/PolysS,用IPTG诱导,进行蛋白表达、纯化。之后将TB10.4,PPD,TB10.4及MPT83混合蛋白作为抗原用间接ELISA法检测牛结核病。结果使用结核杆菌早期分泌蛋白TB10.4作为抗原用间接ELISA方法快速检测牛结核病,并区分BCG免疫的健康牛和感染牛的结果显示灵敏度为23%,没有假阳性反应。结论用PPD作为抗原进行检测灵敏度达到48%,但假阳性率达到21%;用TB10.4与MPT83混合抗原检测时灵敏度下降到6%,假阳性率1%。

引用

页码：932 / 935

页数：4

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