RNA病毒核酸检测质控品大规模生产的研究

Research on the large-scale production of RNA virus nucleic acid control materials

目的大批量、低成本的质控品是核酸检测质量保证中必不可少的原料,针对RNA病毒质控品的大规模制备鲜有研究。方法利用装甲RNA技术,首次通过大肠杆菌发酵技术进行冠状病毒噬菌体病毒样颗粒(假病毒)包装表达。结果通过对大肠杆菌发酵条件进行优化,可稳定制备出单批产能>0.3 L的冠状病毒假病毒。该冠状病毒假病毒包装完整,且线性及稳定性良好,满足样品大规模投料使用需求。具有耐核酸酶、高稳定性、易保存和运输等优点,可被广泛应用于冠状病毒核酸检测试剂盒。结论RNA病毒检测质控品可通过发酵技术实现大规模生产及应用。

Objective Large batch and low-cost quality control products are essential raw materials for quality assurance in nucleic acid testing,and there is little research on the large-scale preparation of RNA virus control materials.Methods Using armored RNA technology,coronavirus phage virus like particles(pseudovirus)were packaged and expressed by Escherichia coli fermentation technology for the first time.Results By optimizing the fermentation conditions of E.coli,a single batch of coronavirus pseudovirus with capacity>0.3L cuold be stably prepared.The coronavirus pseudovirus has complete packaging,good linearity and stability,and can meet the needs of large-scale feeding of samples.It has the advantages of nuclease resistance,high stability,easy storage and transportation,and can be widely used in novel coronavirus nucleic acid test kits.Conclusion RNA virus detection quality control products can be produced and applied on a large scale through fermentation technology.