主要禽类呼吸道病毒诊断技术进展：综述

文献信息

中文摘要

禽类呼吸道疾病，尤其是由禽流感病毒（AIV）、新城疫病毒（NDV）、传染性支气管炎病毒（IBV）和传染性喉气管炎病毒（ILTV）引起的疾病，对全球家禽业构成重大经济威胁。其中数种病毒还可跨物种传播，对公共卫生安全构成风险。鉴于不同病原体引起相似临床症状、共感染流行以及病毒快速突变所带来的挑战，快速、精确且高度灵敏的诊断技术对于有效的疾病预防与控制、疫情监测和流行病学研究至关重要。本综述概述了主要禽类呼吸道病毒的分子和免疫学诊断技术。所有涵盖的技术均按关键性能指标进行评估和比较：靶标类型、分析灵敏度、多重检测能力、周转时间、即时诊断技术（POCT）适用性和应用场景。具体而言，本综述详细介绍了实时定量PCR（qPCR）及其多重形式、等温扩增方法（包括环介导等温扩增（LAMP）、重组酶辅助扩增（RAA）和重组酶聚合酶扩增（RPA）），以及创新的基于CRISPR-Cas的核酸检测系统。本综述还探讨了高通量测序、微阵列技术，以及由纳米材料、重组蛋白和单域抗体增强的先进免疫学检测方法。本综述进一步讨论了集成化、自动化和便携式诊断平台，包括微流控和智能手机辅助系统。最后，本综述概述了禽类诊断技术的未来趋势，为构建更高效、更可及的禽类疾病监测和预警系统提供了实用参考。

英文摘要

Avian respiratory diseases, particularly those induced by the avian influenza virus (AIV), Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and infectious laryngotracheitis virus (ILTV), represent a substantial economic threat to the global poultry industry. Several of these viruses can also transmit across species, posing risks to public health security. Given the challenges posed by similar clinical symptoms from different pathogens, the prevalence of co-infections, and rapid viral mutations, rapid, precise, and highly sensitive diagnostic techniques are imperative for effective disease prevention and control, epidemic surveillance, and epidemiological research. This review provides an overview of molecular and immunological diagnostic techniques for major avian respiratory viruses. All covered technologies are evaluated and compared across key performance metrics: target type, analytical sensitivity, multiplex capacity, turnaround time, point-of-care diagnostic techniques (POCT) suitability and application scenarios. Specifically, the review details real-time quantitative PCR (qPCR) and its multiplex formats, isothermal amplification methods including loop-mediated isothermal amplification (LAMP), recombinase-aided amplification (RAA) and recombinase polymerase amplification (RPA), and innovative CRISPR-Cas-based nucleic acid detection systems. It also examines high-throughput sequencing, microarray technologies, and advanced immunological assays enhanced by nanomaterials, recombinant proteins, and single-domain antibodies. The review further discusses integrated, automated, and portable diagnostic platforms, including microfluidics, and smartphone-assisted systems. Finally, the review outlines future trends in avian diagnostic technologies, providing a practical reference for building more efficient and accessible avian disease surveillance and early warning systems.