基于核糖核酸酶依赖型探针和3D打印芯片的新型冠状病毒变体的快速检测

2019新型冠状病毒（2019-nCoV）蔓延全球之际正伴随着不断演变，已形成三大主要变体毒株B.1.1.7、B.1.351和P.1，能增强传染性和具备逃避疫苗免疫的潜在风险。现阶段，对2019-nCoV变体毒株的检测主要采用过程繁琐的RT-PCR结合产物测序的方法，尚无现场快速检测手段，难以满足防疫工作的时效要求。本项目拟基于核糖核酸酶依赖型探针和3D打印芯片建立对上述三种2019-nCoV变体毒株的快速检测，以适用于现场检测，弥补当前新冠防疫所面临的的技术短缺。核糖核酸酶依赖型探针具有识别单碱基突变能力，与等温核酸扩增技术结合可实现快速、高特异性、高灵敏度、可视化的突变鉴定。同时，利用3D打印技术研制集核酸提取、核酸扩增和产物检测于一体的检测芯片，具备简单、实用、成本低和易产业化等特点。本项目的成功实施有望为监控2019-nCoV变体毒株和应对未来疫情发展提供技术支持。

Novel coronavirus (2019-nCoV) has evolved into three main variants of B.1.1.7, B.1.351, and P.1 during ongoing coronavirus disease 2019 (COVID-19) pandemic, which spread more quickly and have the potential risk of evading vaccine-induced immunity. Presently, time-consuming RT-qPCR assay followed by product sequencing is the main approach of detecting these variants and onsite rapid detection methods are rarely available, hardly meeting rapidness and effectiveness demanded by current epidemic prevention work. This study is going to develop rapid and onsite detection of the variants based on ribonuclease-dependent probe and 3D-printed chip, making up the shortage of technology for COVID-19 prevention. Ribonuclease-dependent probe is able to recognize single nucleotide mutation and realize rapid, highly specific, highly sensitive and visual mutation identification through coupling with isothermal nucleic acid amplification. Meanwhile, a test chip can be fabricated using 3D printing and integrated with nucleic acid extraction, amplification and product detection, possessing the advantages of simplicity, practicability, cost-efficiency and easy commercialization. Our study will provide a technical support for monitoring 2019-nCoV variants and managing future development of COVID-19 pandemic.