非洲马瘟诊断技术研究进展

A Review of Diagnostic Techniques for African Horse Sickness

非洲马瘟(African horse sickness,AHS)是由非洲马瘟病毒(African horse sickness virus,AHSV)引起的一种以发热、皮下水肿及脏器出血为特征的马属动物急性和亚急性虫媒传染病,具有高传染性和快速致病的特点,被世界动物卫生组织列为法定报告疾病,中国也将其列为一类动物疫病。世界动物健康信息系统数据显示,AHS近几年在非洲和与中国邻近的东南亚地区流行,形势非常严峻,严格监测边境地区和口岸的马属动物AHS,及时清除带毒动物是中国AHS防控的法定策略。笔者对AHS的病原学和血清学检测技术研究进展进行了整理和总结。病原学诊断技术主要包括病毒分离、RT-PCR、恒温扩增和基因芯片技术等,其中病毒分离是经典的确诊方法,RT-PCR和实时荧光定量RT-PCR技术为常用的实验室诊断方法,恒温扩增技术如环介导等温扩增技术(LAMP)、重组酶聚合酶扩增技术(RPA)和重组酶介导的扩增技术(RAA)等可用于现场快速诊断,基因芯片技术可实现多病原检测。血清学诊断包括ELISA、病毒中和试验和补体结合试验等。其中RT-PCR、实时荧光定量RT-PCR和ELISA均为世界动物卫生组织和《非洲马瘟诊断技术》(GB/T 21675—2022)推荐的主要检测方法。近些年来,基于高通量测序的宏基因组技术能同时对多种病原体进行定量检测,极大提高了疾病诊断的准确性和效率;基于云平台和物联网的疾病智能监测和预警系统等新技术的发展与应用,将为AHS的监测和预防提供更为科学和精确的技术支持。

African horse sickness(AHS)is an acute and subacute insect borne infectious and highly contagious and rapidly fatal infectious disease of equids caused by African horse sickness virus(AHSV).It is characterized by symptoms such as fever,subcutaneous swelling,and internal organ hemorrhaging.AHS is classified as a notifiable disease by the World Animal Health Organization and is also categorized as ClassⅠanimal diseases in China.Recent data from the World Animal Health Information System indicates that AHS has been spreading in Africa and neighboring Southeast Asian regions,which are in close proximity to China.The situation is considered extremely severe.China’s official strategy for the prevention and control of AHS involves strict monitoring of equids with a focus on border areas and ports,along with the timely removal of infected animals.This article provides a comprehensive overview of the advancements and recent research developments in the field of AHS diagnostics,both in terms of pathogenetics and serology.Pathogenetic diagnostic techniques encompass virus isolation,RT-PCR,isothermal amplification,and gene chip technology.Virus isolation remains the classical gold standard for definitive diagnosis.RT-PCR and Real-time quantitative RT-PCR techniques are commonly employed in laboratory settings.Isothermal amplification methods such as loop-mediated isothermal amplification(LAMP),recombinase polymerase amplification(RPA),and recombinase-aided amplification(RAA)are suited for rapid on-site diagnosis.Gene chip technology allows for the simultaneous detection of multiple pathogens.In the realm of serological diagnostics,methods include ELISA,virus neutralization tests,and complement fixation tests.Among these,RT-PCR,Real-time quantitative RT-PCR,and ELISA are recommended as the primary diagnostic methods by both the World Animal Health Organization and the“African Horse Sickness Diagnostic Techniques”(GB/T 21675—2022)guidelines.In recent years,the emergence of high-throughput metagenomic techniques based on next-generation sequencing has enabled simultaneous quantitative detection of various pathogens,greatly enhancing diagnostic accuracy and efficiency of dieases.The development and application of new technologies,such as cloud-based platforms and the Internet of Things,in disease intelligence monitoring and warning systems,promise to offer more scientifically rigorous and precise technical support for the monitoring and prevention of AHS.