tetano
Editor, Senior Moderator
Sens Actuators B Chem
. 2021 Oct 15;345:130411.
doi: 10.1016/j.snb.2021.130411. Epub 2021 Jul 6.
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
Yaqin Zhang[SUP] 1 [/SUP], Minyan Chen[SUP] 2 [/SUP], Chengrong Liu[SUP] 3 [/SUP], Jiaqi Chen[SUP] 2 [/SUP], Xinyi Luo[SUP] 4 [/SUP], Yingying Xue[SUP] 4 [/SUP], Qiming Liang[SUP] 3 5 6 [/SUP], Li Zhou[SUP] 7 [/SUP], Yu Tao[SUP] 8 [/SUP], Mingqiang Li[SUP] 8 [/SUP], Di Wang[SUP] 1 2 [/SUP], Jianhua Zhou[SUP] 4 [/SUP], Jiasi Wang[SUP] 4 [/SUP]
Affiliations
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.
Keywords: AuNP, gold nanoparticle; COVID-19, Corona Virus Disease 2019; CRISPR, clustered regularly interspaced short palindromic repeats; CRISPR/Cas; Coronavirus disease; DMEM, Dulbecco’s modified Eagle’s medium; FDA, American Food and Drug Administration; Gold nanoparticle; HCRs, hybridization chain reactions; High-throughput on-site detection; LAMP, loop-mediated isothermal amplification; Loop-mediated isothermal amplification; NMPA, the Chinese National Medical Products Administration; POCT, point of care testing; RPA, recombinase polymerase amplification; RT-qPCR, reverse transcription-real time quantitative PCR; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; TCEP, Tris(2-carboxyethyl) phosphine; TEM, transmission electron microscopy.
. 2021 Oct 15;345:130411.
doi: 10.1016/j.snb.2021.130411. Epub 2021 Jul 6.
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
Yaqin Zhang[SUP] 1 [/SUP], Minyan Chen[SUP] 2 [/SUP], Chengrong Liu[SUP] 3 [/SUP], Jiaqi Chen[SUP] 2 [/SUP], Xinyi Luo[SUP] 4 [/SUP], Yingying Xue[SUP] 4 [/SUP], Qiming Liang[SUP] 3 5 6 [/SUP], Li Zhou[SUP] 7 [/SUP], Yu Tao[SUP] 8 [/SUP], Mingqiang Li[SUP] 8 [/SUP], Di Wang[SUP] 1 2 [/SUP], Jianhua Zhou[SUP] 4 [/SUP], Jiasi Wang[SUP] 4 [/SUP]
Affiliations
- PMID: 34248284
- PMCID: PMC8257267
- DOI: 10.1016/j.snb.2021.130411
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.
Keywords: AuNP, gold nanoparticle; COVID-19, Corona Virus Disease 2019; CRISPR, clustered regularly interspaced short palindromic repeats; CRISPR/Cas; Coronavirus disease; DMEM, Dulbecco’s modified Eagle’s medium; FDA, American Food and Drug Administration; Gold nanoparticle; HCRs, hybridization chain reactions; High-throughput on-site detection; LAMP, loop-mediated isothermal amplification; Loop-mediated isothermal amplification; NMPA, the Chinese National Medical Products Administration; POCT, point of care testing; RPA, recombinase polymerase amplification; RT-qPCR, reverse transcription-real time quantitative PCR; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; TCEP, Tris(2-carboxyethyl) phosphine; TEM, transmission electron microscopy.