Pyrococcus furiosus Argonaute coupled with modified ligase chain reaction for detection of SARS-CoV-2 and HPV

被引:43
作者
Wang, Longyu [1 ]
He, Ruyi [1 ]
Lv, Bin [1 ]
Yu, Xiao [2 ]
Liu, Yang [1 ]
Yang, Jun [1 ]
Li, Wenqiang [1 ]
Wang, Yuan [3 ]
Zhang, Hang [1 ]
Yan, Guangbo [1 ]
Mao, Wuxiang [1 ]
Liu, Linlin [2 ]
Wang, Fei [1 ]
Ma, Lixin [1 ]
机构
[1] Hubei Univ, Hubei Collaborat Innovat Ctr Green Transformat Bi, Sch Life Sci, Hubei Key Lab Ind Biotechnol,State Key Lab Biocat, Wuhan, Hubei, Peoples R China
[2] Hubei Prov Ctr Dis Control & Prevent, Wuhan, Hubei, Peoples R China
[3] Hubei Univ Arts & Sci, Med Coll, Xiangyang, Hubei, Peoples R China
基金
中国国家自然科学基金;
关键词
Prokaryotic argonaute; Ligase chain reaction; SARS-CoV-2; HPV; Nucleic acid detection; NUCLEIC-ACID DETECTION; LIGATION; QUANTIFICATION; MICRORNA;
D O I
10.1016/j.talanta.2021.122154
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Infectious diseases caused by viruses such as SARS-CoV-2 and HPV have greatly endangered human health. The nucleic acid detection is essential for the early diagnosis of diseases. Here, we propose a method called PLCR (PfAgo coupled with modified Ligase Chain Reaction for nucleic acid detection) which utilizes PfAgo to only use DNA guides longer than 14-mer to specifically cleave DNA and LCR to precisely distinguish single-base mismatch. PLCR can detect DNA or RNA without PCR at attomolar sensitivities, distinguish single base mutation between the genome of wild type SARS-CoV-2 and its mutant spike D614G, effectively distinguish the novel coronavirus from other coronaviruses and finally achieve multiplexed detection in 70 min. Additionally, LCR products can be directly used as DNA guides without additional input guides to simplify primer design. With desirable sensitivity, specificity and simplicity, the method can be extended for detecting other pathogenic microorganisms.
引用
收藏
页数:8
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