An integrated dual-layer microfluidic platform for multiple respiratory viruses screening

Hua Wang; Jingsong Xu; Shun Li; Xi Wang; Gang Liu; Shuang Yang; Fei Zhao; Qian Liu; Xiang Chen; Chao He; Min Li

doi:10.1016/j.aca.2023.340812

An integrated dual-layer microfluidic platform for multiple respiratory viruses screening

Anal Chim Acta. 2023 Feb 15:1242:340812. doi: 10.1016/j.aca.2023.340812. Epub 2023 Jan 7.

Authors

Hua Wang¹, Jingsong Xu², Shun Li³, Xi Wang², Gang Liu⁴, Shuang Yang², Fei Zhao⁵, Qian Liu², Xiang Chen⁶, Chao He⁷, Min Li⁸

Affiliations

¹ Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China. Electronic address: fdeduwh@126.com.
² Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
³ Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
⁴ Bio Metrology Laboratory, Chemical and Ionizing Radiation Metrolology Institute, Shanghai Institute of Measurement and Testing Technology, Shanghai, China.
⁵ National Key Laboratory of Science and Technology on Micro/Nano Fabrication, Department of Micro/Nano Electronics, Shanghai Jiao Tong University, Shanghai 200240, China.
⁶ National Key Laboratory of Science and Technology on Micro/Nano Fabrication, Department of Micro/Nano Electronics, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: xiangchen@sjtu.edu.cn.
⁷ Department of Laboratory Medicine, West China Hospital of Sichuan University, Sichuan, China. Electronic address: hc_wsw@126.com.
⁸ Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China. Electronic address: ruth_limin@126.com.

Abstract

Currently, the coronavirus disease 2019 (COVID-19) caused by the outbreak of a novel coronavirus (SARS-CoV-2) is spreading rapidly worldwide. Due to the high incidence of influenza coinciding with SARS-CoV-2, rapid detection is crucial to prevent spreading. Here, we present an integrated dual-layer microfluidic platform for specific and highly sensitive SARS-CoV-2, influenza viruses A (FluA) H1N1, H3N2, and influenza virus B (FluB) simultaneous detection. The platform includes a dual microchip (Dμchip) and a portable detection device for real-time fluorescence detection, temperature control and online communication. The Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) and Cas12a cleavage were performed on the Dμchip. The limit of detection (LoD) of the Dμchip assay was 10 copies for SARS-CoV-2, FluA H1N1, H3N2, and FluB RNAs. The Dμchip assay yielded no cross-reactivity against other coronaviruses, so it was suitable for the screening of multiple viruses. Moreover, the positive percentage agreement (PPA) and negative percentage agreement (NPA) of the assay were 97.9% and 100%, respectively, in 75 clinical samples compared to data from RT-PCR-based assays. Furthermore, the assay allowed the detection SARS-CoV-2 and influenza viruses in spiked samples. Overall, the present platform would provide a rapid method for the screening of multiple viruses in hospital emergency, community and primary care settings and facilitate the remote diagnosis and outbreak control of the COVID-19.

Keywords: CRISPR/Cas12a; Dual microchip; Influenza viruses; SARS-CoV-2.

MeSH terms

COVID-19* / diagnosis
Humans
Influenza A Virus, H1N1 Subtype* / genetics
Influenza A Virus, H3N2 Subtype / genetics
Microfluidics
Nucleic Acid Amplification Techniques / methods
RNA, Viral
SARS-CoV-2
Sensitivity and Specificity

Substances

RNA, Viral