A fully automated centrifugal microfluidic system for sample-to-answer viral nucleic acid testing
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tps://doi.org/10.1007/s11426-020-9800-6
SPECIAL ISSUE: 2020 Emerging Investigator Issue
A fully automated centrifugal microfluidic system for sample-to-answer viral nucleic acid testing Fei Tian
1,2†
1,2†
1,2
1,2
3*
, Chao Liu , Jinqi Deng , Ziwei Han , Lu Zhang , 1* 1,2* Qinghua Chen & Jiashu Sun
1
CAS Key Laboratory of Standardization and Measurement of Nanotechnology, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology, Beijing 100190, China; 2 University of Chinese Academy of Sciences, Beijing 100049, China; 3 Department of Chemistry, Capital Normal University, Beijing 100048, China Received May 7, 2020; accepted June 23, 2020; published online July 27, 2020
The outbreak of virus-induced infectious diseases poses a global public-health challenge. Nucleic acid amplification testing (NAAT) enables early detection of pandemic viruses and plays a vital role in preventing onward transmission. However, the requirement of skilled operators, expensive instrumentation, and biosafety laboratories has hindered the use of NAAT for screening and diagnosis of suspected patients. Here we report development of a fully automated centrifugal microfluidic system with sample-in-answer-out capability for sensitive, specific, and rapid viral nucleic acid testing. The release of nucleic acids and the subsequent reverse transcription loop-mediated isothermal amplification (RT-LAMP) were integrated into the reaction units of a microfluidic disc. The whole processing steps such as injection of reagents, fluid actuation by rotation, heating and temperature control, and detection of fluorescence signals were carried out automatically by a customized instrument. We validate the centrifugal microfluidic system using oropharyngeal swab samples spiked with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) armored RNA particles. The estimated limit of detection for armored RNA particles is 2 copies per reaction, the throughput is 21 reactions per disc, and the assay sample-to-answer time is approximately 70 min. This enclosed and automated microfluidic system efficiently avoids viral contamination of aerosol, and can be readily adapted for virus detection outside the diagnostic laboratory. nucleic acid testing, virus, microfluidics, sample-to-answer, automation Citation:
1
Tian F, Liu C, Deng J, Han Z, Zhang L, Chen Q, Sun J. A fully automated centrifugal microfluidic system for sample-to-answer viral nucleic acid testing. Sci China Chem, 2020, 63, https://doi.org/10.1007/s11426-020-9800-6
Introduction
Infectious diseases caused by pathogens are one of the leading causes of death in the world. According to the statistics provided by the world health organization (WHO), the number of deaths attributable to pathogen infections such as hepatitis, tuberculosis and malaria is 4.3 million worldwide
†These authors contributed equally to this work. *Corresponding authors (email: [email protected]; [email protected]; [email protected])
in 2016 [1]. The emergence and spread of pathogens such
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