1. Sensitive and rapid on-site detection of SARS-CoV-2 using a gold nanoparticle-based high-throughput platform coupled with CRISPR/Cas12-assisted RT-LAMP
- Author
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Yaqin Zhang, Yingying Xue, Li Zhou, Xinyi Luo, Di Wang, Jiaqi Chen, Jianhua Zhou, Chengrong Liu, Qiming Liang, Yu Tao, Jiasi Wang, Minyan Chen, and Mingqiang Li
- Subjects
Gold nanoparticle ,Coronavirus disease 2019 (COVID-19) ,Computer science ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,RPA, recombinase polymerase amplification ,NMPA, the Chinese National Medical Products Administration ,Loop-mediated isothermal amplification ,AuNP, gold nanoparticle ,COVID-19, Corona Virus Disease 2019 ,Computational biology ,Virus diseases ,LAMP, loop-mediated isothermal amplification ,SARS-CoV-2, severe acute respiratory syndrome coronavirus 2 ,Article ,DMEM, Dulbecco’s modified Eagle’s medium ,CRISPR/Cas ,HCRs, hybridization chain reactions ,Materials Chemistry ,POCT, point of care testing ,CRISPR ,TCEP, Tris(2-carboxyethyl) phosphine ,Electrical and Electronic Engineering ,TEM, transmission electron microscopy ,Instrumentation ,Throughput (business) ,ComputingMethodologies_COMPUTERGRAPHICS ,FDA, American Food and Drug Administration ,High-throughput on-site detection ,Metals and Alloys ,Condensed Matter Physics ,Coronavirus disease ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials ,Microplate Reader ,CRISPR, clustered regularly interspaced short palindromic repeats ,Population screening ,RT-qPCR, reverse transcription-real time quantitative PCR - Abstract
Graphical abstract, The outbreak of corona virus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to a global pandemic. The high infectivity of SARS-CoV-2 highlights the need for sensitive, rapid and on-site diagnostic assays of SARS-CoV-2 with high-throughput testing capability for large-scale population screening. The current detection methods in clinical application need to operate in centralized labs. Though some on-site detection methods have been developed, few tests could be performed for high-throughput analysis. We here developed a gold nanoparticle-based visual assay that combines with CRISPR/Cas12a-assisted RT-LAMP, which is called Cas12a-assisted RT-LAMP/AuNP (CLAP) assay for rapid and sensitive detection of SARS-CoV-2. In optimal condition, we could detect down to 4 copies/μL of SARS-CoV-2 RNA in 40 min. by naked eye. The sequence-specific recognition character of CRISPR/Cas12a enables CLAP a superior specificity. More importantly, the CLAP is easy for operation that can be extended to high-throughput test by using a common microplate reader. The CLAP assay holds a great potential to be applied in airports, railway stations, or low-resource settings for screening of suspected people. To the best of our knowledge, this is the first AuNP-based colorimetric assay coupled with Cas12 and RT-LAMP for on-site diagnosis of COVID-19. We expect CLAP assay will improve the current COVID-19 screening efforts, and make contribution for control and mitigation of the pandemic.
- Published
- 2021