摘要
利用分子克隆技术,提取结核分支杆菌早期分泌性抗原靶ESAT-6基因,然后对其进行克隆表达,获得高效表达的ESAT-6蛋白,Western blot证实ESAT-6蛋白可与结核牛阳性血清发生特异性反应。以纯化复性的ESAT-6重组蛋白作为诊断抗原建立的间接ELISA特异性为94%、敏感性为63.35%。交叉反应试验表明,该抗原只与牛副结核病阳性血清有交叉反应,而不与其他5种常见的牛病阳性血清发生交叉反应。用间接ELISA方法对285份PPD阳性牛血清、对74份PPD阴性牛血清、79份牛γ-干扰素(IFN-γ)检测阳性血清进行检测,其负荷率分别为62.1%、98.64%、83.54%。实验结果表明建立的间接ELISA方法可以用于牛结核病感染和免疫抗体检测,且具有很好的开发和应用前景。
To study cloning,identification and expression of ESAT 一 6 gene from Mycobacterium tuberculosis for the basis of development of diagnostic method of tuberculosis.Purified ESAT-6 protein reacted positively with serotype-specific cattle sera of bovine tuberculosis by Western blot.An indirect ELISA（ESAT-6-ELISA） was developed using purified protein as coating antigen to detect bovine tuberculosis antibodies in cattle.Test of serum samples from cows with bovine tuberculosis and serum samples from healthy cattle detected 63.35% positive with a specificity of 94.0%.The assay was highly specific and showed no crossreaction with the positive sera of other bovine diseases.Test of 285 samples from bovine with positive skin test of PPD,74 samples from bovine with negative bovine skin test of PPD and 79 samples from bovine with positive test of IFN-γshowed a coincidence rate of 62.1%,98.64% and 83.54% against cattle serum.An indirect ELISA assay was established by using ESAT-6 protein as coating antigen for the antibody detection against bovine tuberculosis.
出处
《草食家畜》
2010年第2期69-72,共4页
Grass-Feeding Livestock
基金
新疆少数民族科技人才特殊培养计划科研项目"基于基因重组蛋白的牛结核病ELISA诊断方法的研究"
