Dual-target nucleic acid sequences responsive electrochemiluminescence biosensor using single type carbon dots as probe for SARS-CoV-2 detection based on series catalytic hairpin assembly amplification

Ying Zhang; Xiaocui Huang; Weixin Li; Qunfang Xie; Jie Zhang; Fang Luo; Bin Qiu; Zhonghui Chen; Zhenyu Lin; Guoyan Xu

doi:10.1016/j.snb.2022.133223

Dual-target nucleic acid sequences responsive electrochemiluminescence biosensor using single type carbon dots as probe for SARS-CoV-2 detection based on series catalytic hairpin assembly amplification

Sens Actuators B Chem. 2023 Mar 15:379:133223. doi: 10.1016/j.snb.2022.133223. Epub 2022 Dec 22.

Authors

Ying Zhang¹, Xiaocui Huang², Weixin Li², Qunfang Xie³, Jie Zhang⁴, Fang Luo², Bin Qiu², Zhonghui Chen⁵, Zhenyu Lin², Guoyan Xu³

Affiliations

¹ Central Laboratory, Fujian Key Laboratory of Precision Medicine for Cancer, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, China.
² Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China.
³ Department of General Practice, The Center of Health Management, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, China.
⁴ Department of Geriatrics, Branch of National Clinical Research Center for Aging and Medicine, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, China.
⁵ Affiliated Hospital of Putian University, Putian University, Putian, Fujian 351100, China.

Abstract

Severe acute respiratory syndrome coronavirus (SARS-CoV-2) is rampant all over the world, and rapid and effective virus detection is the best auxiliary to curb the spread of the epidemic. A diagnosis can only be made if two or more different nucleic acid sequences are confirmed at the same time, and in most of traditional detection technologies, these target sequences have been detected separately. In this work, an electrochemiluminescent (ECL) biosensor employing a single ECL probe as signal output and responding to dual-target simultaneously is proposed for the first time. Taking the two sequences located in ORF 1ab region and N region of SARS-CoV-2 gene sequence as the model target and nitrogen doped carbon quantum dots (CDs) as ECL beacon, supplemented with catalytic hairpin assembly (CHA) reaction for signal amplification, the presented strategy has been successfully applied to the rapid detection of SARS-CoV-2. The developed SARS-CoV-2 biosensor based on the series CHA systems can realize the quantitative determination of SARS-CoV-2 in the range of 50 fM to 200 pM within 40 min. Moreover, the clinical validity of this method has been verified by the high consistency between the detection results of using this method and those using RT-qPCR for seven clinical pharyngeal swab samples.

Keywords: Carbon quantum dots; Catalytic hairpin assembly; Dual-target detection; Electrochemiluminescence; SARS-CoV-2.