核酸
核糖核酸
多路复用
化学
病毒学
滚动圆复制
计算生物学
多重位移放大
分子生物学
生物
聚合酶链反应
聚合酶
DNA
基因
遗传学
生物化学
DNA提取
作者
Fukai Wang,Xiaochen Ma,Jing Ye,Chunhong Shi,Yun Cui,Zhongqi Yu,Tianming Li,Donglei Yang,Min Li,Pengfei Wang
标识
DOI:10.1021/acs.analchem.3c03548
摘要
Detection of viral infections (e.g., SARS-CoV-2) with high precision is critical to disease control and treatment. There is an urgent need to develop point-of-care detection methods to complement the gold standard laboratory-based PCR assay with comparable sensitivity and specificity. Herein, we developed a method termed mCAD to achieve ultraspecific point-of-care detection of SARS-CoV-2 RNA while maintaining high sensitivity by programming multiplex rolling circle amplification and toehold-mediated strand displacement reactions. RCA offers sufficient amplification of RNA targets for subsequent detection. Most importantly, a multilayer of detection specificity is implemented into mCAD via sequence-specific hybridization of nucleic acids across serial steps of this protocol to fully eliminate potential false-positive detections. Using mCAD, we demonstrated a highly specific, sensitive, and convenient visual detection of SARS-CoV-2 RNA from both synthetic and clinical samples, exhibiting performance comparable to qPCR. We envision that mCAD will find its broad applications in clinical prospects for nucleic acid detections readily beyond SARS-CoV-2 RNA.
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