猪瘟病毒装甲RNA颗粒质控品的研制及评估

Development and evaluation of quality control products for swine fever virus armored RNA particles

[目的]研制一种安全稳定高效的猪瘟核酸检测质控品,为猪瘟疫病监测提供技术支持。[方法]将猪瘟病毒核酸检测靶基因构入至新型装甲RNA表达载体pTMACC,转化、表达并纯化,制备猪瘟病毒装甲RNA质控品。借助电泳、电镜、动态散射分析鉴定该物质结构特性,并对其均匀性、稳定性进行分析,评估其实用性能。[结果]该装甲RNA颗粒含有猪瘟病毒特异性基因,序列溯源至法国Thiverval株;为RNA-蛋白复合物,颗粒呈直径约26 nm的多边形;该质控品均匀、稳定,-20℃至少稳定保存36个月;经8家权威实验室性能评估,该物质可以作为猪瘟病毒核酸检测质控品。[结论]猪瘟病毒装甲RNA质控品均匀性良好,稳定性强,无生物传染性,核酸RNA不易降解,可稳定保存,进一步保障猪瘟疫病的临床检测的准确性。

[Objectives]To develop a safe,stable and efficient quality control product for nucleic acid test of swine fever,and provide technical support for the monitoring of swine plague.[Methods]The target gene of CSFV nucleic acid test was constructed into a novel armored RNA expression vector pTMACC.The pTMACC CSFV recombinant plasmid was transformed,expressed,purified to prepare the quality control product for swine fever virus armored RNA particles.The structural characteristics of the armored RNA particles were identified with electrophoresis,electron microscopy,and dynamic scattering analysis.Its uniformity and stability were analyzed,and its clinical performance was evaluated.[Results]The armored RNA particle contained a specific gene of the swine fever virus,and its sequence was traced back to the Thiverval strain in France.It was an RNA-protein complex with polygonal particles with a diameter of approximately 26 nm.The quality control product is uniform and stable,and should be stored at-20℃for at least 36 months.According to the performance evaluation of 8 authoritative laboratories,the armored RNA particle can be used as the quality control product for the nucleic acid test of CSFV.[Conclusions]The armored RNA particle as quality control product of swine fever virus has good uniformity,strong stability,and no biological infectivity.The nucleic acid RNA is not easily degraded and can be stably stored,further ensuring the accuracy of clinical detection of swine fever disease.