摘 要
牛结核病作为一种重要的人畜共患病，严重威胁畜牧业发展和公共卫生安全。本研究针对传统检测方法耗时长、灵敏度低等问题，开发了一种基于重组蛋白和纳米材料的快速检测技术。研究首先通过生物信息学分析筛选出特异性抗原MPB70和MPB83，采用原核表达系统进行重组表达并纯化；随后利用量子点标记技术构建荧光免疫层析试纸条，优化了反应体系参数。为验证该技术的有效性，采集了来自5个规模化牧场的1200份牛血清样本，采用双盲法与传统ELISA方法进行对比测试。实验结果表明，所开发的快速检测技术在15分钟内即可完成检测，灵敏度达到95.3%，特异性为97.8%，与ELISA方法的符合率为96.5%。进一步研究表明，该方法对早期感染病例的检出率较传统方法提高23.6%，且不受常见干扰物质影响。现场应用评估显示，该技术操作简便、结果直观，适合基层兽医站和养殖场使用。
关键词：牛结核病，快速检测技术，量子点标记，重组蛋白
Abstract
As an important zoonotic disease, bovine tuberculosis poses a serious threat to animal husbandry development and public health security. In this study, a rapid detection technology based on recombinant protein and nanomaterials was developed to solve the problems of long time and low sensitivity of traditional detection methods. In this study, the specific antigens MPB70 and MPB83 were screened by bioinformatics analysis, and recombinant ex pression and purification were performed by prokaryotic ex pression system. Then the fluorescence immunochromatographic test strip was constructed using quantum dot labeling technology, and the parameters of the reaction system were optimized. To verify the effectiveness of this technique, 1200 bovine serum samples from 5 large-scale farms were collected and compared with traditional ELISA method by double-blind method. The results show that the developed rapid detection technology can be completed within 15 minutes, the sensitivity is 95.3%, the specificity is 97.8%, and the coincidence rate with the ELISA method is 96.5%. Further study showed that the detection rate of early infection cases was 23.6% higher than that of traditional methods, and it was not affected by common interfering substances. Field application evaluation shows that the technology is easy to operate, intuitive results, suitable for basic veterinary stations and farms.
Key words: Bovine tuberculosis, rapid detection technology, quantum dot labeling, recombinant protein
目 录
1 绪论 1
1.1 牛结核病快速检测技术研究背景 1
1.2 牛结核病快速检测技术研究意义 1
1.3 牛结核病快速检测技术研究现状 2
2 牛结核病快速检测技术开发 2
2.1 现有检测技术的局限性分析 2
2.2 新型快速检测技术原理设计 3
2.3 关键技术参数优化研究 4
2.4 检测试剂盒的研制与验证 4
3 牛结核病快速检测技术应用评估 5
3.1 现场应用试验方案设计 5
3.2 不同养殖环境下的适用性评估 6
3.3 与传统检测方法的对比分析 6
3.4 经济效益与社会效益评估 7
4 牛结核病快速检测技术推广策略 8
4.1 技术推广面临的挑战分析 8
4.2 政策支持与标准制定建议 8
4.3 技术人员培训体系建设 9
4.4 推广应用效果监测机制 10
5 结论 10
参考文献 12
致谢 13
