科研动态

狂犬病毒、伪狂犬病毒、犬瘟热病毒、犬细小病毒和犬冠状病毒是引起犬科动物重要传染病的5种病毒，严重威胁着犬科动物的健康，是出入境犬科动物重点筛查的致病因子。为提升口岸进出境犬科动物检疫工作效率，建立了上述5种病毒微流控芯片检测方法。在建立5种病毒环等温扩增检测技术（loop mediated isothermal amplification，LAMP）的基础上，将这些LAMP检测体系固化在同一块塑料芯片上，制备了同步高通量快速检测微流控芯片，并将其应用于临床检测。结果显示：微流控芯片检测方法具有良好的特异性，含有目的基因的阳性样本仅在芯片上相应病毒反应槽出现显著扩增，而其他病毒反应槽未出现扩增，彼此无交叉反应；微流控芯片的敏感性与LAMP方法一致；通过检测不同时期收集的感染犬瘟热病毒、犬细小病毒和犬冠状病毒临床样本，证实建立的微流控芯片检测方法具有良好的稳定性和可靠性。与现有核酸检测方法相比，微流控芯片可以在受试核酸上样40 min内完成上述5种犬病毒的快速筛查，从而满足进出境犬科动物快速检疫的需求。

Establishment and Application of the Microfluidic Chip Assay for Five Kinds of Major Canine Viruses 

Rabies virus，pseudorabies virus，canine distemper virus，canine parvovirus and canine coronavirus are five kinds of viruses that cause important infectious diseases of canine，which seriously threaten the health of canine，and are the key pathogenic factors to be screened and examined by the exit-entry ports. In order to improve the efficiency of port entry-exit quarantine for canine，a microfluidic chip assay for the above five viruses was established. Five loop-mediated isothermal amplification（LAMP）assays were specifically established and then solidified on the same plastic chip to prepare a microfluidic chip with high-throughput and rapid detection capacity for clinical detection. The results showed that the established assay had a good specificity where positive samples with targeted genes were obviously amplified only in the reaction tank in corresponding viruses instead of other viruses，and no cross reaction with each other was found；the sensitivity of microfluidic chip was consistent with that of LAMP；the established assay was with good stability and reliability as verified through detecting clinical samples infected with canine distemper virus and canine parvovirus and collected in different periods. Compared to existing nucleic acid detection methods，the microfluidic chip could be used to complete rapid screening of the above five viruses within 40 minutes after the loading of nucleic acids，which could satisfy the needs of rapid quarantine for entry-exit canine animals.

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