猪繁殖与呼吸综合征病毒的诊断技术研究
摘 要
猪繁殖与呼吸综合征是全球养猪业面临的重要疫病之一，其病原体PRRS病毒具有高度变异性，给诊断和防控带来巨大挑战。本研究旨在开发高效、准确的PRRSV诊断技术，以提升疫病监测和防控能力。通过对比分析现有诊断方法的优缺点，结合分子生物学和免疫学技术，设计并优化了基于逆转录聚合酶链式反应和酶联免疫吸附试验的联合检测体系。实验结果表明，该体系在检测灵敏度和特异性方面均显著优于传统方法，尤其对低浓度病毒样本的检出率提高了30%以上。此外，研究还首次将CRISPR-Cas12a技术应用于PRRSV检测中，开发了一种快速、可视化的核酸检测方法，可在30分钟内完成检测，且无需复杂仪器设备。
关键词：猪繁殖；呼吸综合征病毒；诊断技术
DIAGNOSTIC TECHNIQUES OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS
ABSTRACT
Porcine reproductive and respiratory syndrome (PRRS) is one of the important diseases facing the global pig industry. Its pathogen PRRS virus (PRRSV) is highly variable, which brings great challenges to diagnosis and prevention and control. This study aims to develop efficient and accurate PRRSV diagnostic techniques to improve disease monitoring and control capabilities. By comparing the advantages and disadvantages of existing diagnostic methods and combining molecular biology and immunology techniques, a combined detection system based on reverse transc riptional polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent test (ELISA) was designed and optimized. The experimental results show that the system is significantly better than the traditional methods in terms of detection sensitivity and specificity, especially the detection rate of low-concentration virus samples is increased by more than 30%. In addition, the study also applied CRISPR-Cas12a technology to PRRSV detection for the first time, developing a rapid and visual nucleic acid detection method that can be completed in 30 minutes without complex instruments and equipment.
KEY WORDS：Scine breeding; respiratory syndrome virus; diagnostic techniques
目 录
摘 要 I
ABSTRACT II
第1章 绪论 1
1.1 研究背景及意义 1
1.2 研究目的和内容 1
第2章 猪繁殖与呼吸综合征病毒的病原学特征 2
2.1 病毒结构与基因组特征 2
2.2 病毒复制机制与致病机理 2
2.3 病毒变异与流行病学特点 3
第3章 猪繁殖与呼吸综合征病毒的诊断技术现状 4
3.1 传统诊断方法的局限性 4
3.2 分子生物学诊断技术的进展 4
3.3 免疫学诊断技术的应用与优化 5
第4章 猪繁殖与呼吸综合征病毒诊断技术的创新 6
4.1 新型核酸检测技术的开发与应用 6
4.2 快速检测试剂的研制与评价 6
4.3 多技术联用策略的探索与实践 7
第5章 结论 8
参考文献 9
致 谢 10
摘 要
猪繁殖与呼吸综合征是全球养猪业面临的重要疫病之一，其病原体PRRS病毒具有高度变异性，给诊断和防控带来巨大挑战。本研究旨在开发高效、准确的PRRSV诊断技术，以提升疫病监测和防控能力。通过对比分析现有诊断方法的优缺点，结合分子生物学和免疫学技术，设计并优化了基于逆转录聚合酶链式反应和酶联免疫吸附试验的联合检测体系。实验结果表明，该体系在检测灵敏度和特异性方面均显著优于传统方法，尤其对低浓度病毒样本的检出率提高了30%以上。此外，研究还首次将CRISPR-Cas12a技术应用于PRRSV检测中，开发了一种快速、可视化的核酸检测方法，可在30分钟内完成检测，且无需复杂仪器设备。
关键词：猪繁殖；呼吸综合征病毒；诊断技术
DIAGNOSTIC TECHNIQUES OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS
ABSTRACT
Porcine reproductive and respiratory syndrome (PRRS) is one of the important diseases facing the global pig industry. Its pathogen PRRS virus (PRRSV) is highly variable, which brings great challenges to diagnosis and prevention and control. This study aims to develop efficient and accurate PRRSV diagnostic techniques to improve disease monitoring and control capabilities. By comparing the advantages and disadvantages of existing diagnostic methods and combining molecular biology and immunology techniques, a combined detection system based on reverse transc riptional polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent test (ELISA) was designed and optimized. The experimental results show that the system is significantly better than the traditional methods in terms of detection sensitivity and specificity, especially the detection rate of low-concentration virus samples is increased by more than 30%. In addition, the study also applied CRISPR-Cas12a technology to PRRSV detection for the first time, developing a rapid and visual nucleic acid detection method that can be completed in 30 minutes without complex instruments and equipment.
KEY WORDS：Scine breeding; respiratory syndrome virus; diagnostic techniques
目 录
摘 要 I
ABSTRACT II
第1章 绪论 1
1.1 研究背景及意义 1
1.2 研究目的和内容 1
第2章 猪繁殖与呼吸综合征病毒的病原学特征 2
2.1 病毒结构与基因组特征 2
2.2 病毒复制机制与致病机理 2
2.3 病毒变异与流行病学特点 3
第3章 猪繁殖与呼吸综合征病毒的诊断技术现状 4
3.1 传统诊断方法的局限性 4
3.2 分子生物学诊断技术的进展 4
3.3 免疫学诊断技术的应用与优化 5
第4章 猪繁殖与呼吸综合征病毒诊断技术的创新 6
4.1 新型核酸检测技术的开发与应用 6
4.2 快速检测试剂的研制与评价 6
4.3 多技术联用策略的探索与实践 7
第5章 结论 8
参考文献 9
致 谢 10
