tetano
Editor, Senior Moderator
Biosens Bioelectron
. 2021 Dec 8;199:113865.
doi: 10.1016/j.bios.2021.113865. Online ahead of print.
Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system
Ziyue Li[SUP] 1 [/SUP], Xiong Ding[SUP] 2 [/SUP], Kun Yin[SUP] 2 [/SUP], Lori Avery[SUP] 3 [/SUP], Enrique Ballesteros[SUP] 3 [/SUP], Changchun Liu[SUP] 4 [/SUP]
Affiliations
Abstract
Rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for early diagnostics and timely medical treatment of coronavirus disease 2019 (COVID-19). However, current detection methods typically rely on expensive and bulky instrumentation. Here, we developed a simple, sensitive, instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using a self-contained microfluidic system. The microfluidic chip integrates isothermal amplification, CRISPR cleavage, and lateral flow detection in a single, closed microfluidic platform, enabling contamination-free, visual detection. To simplify the operation and transportation of the device, we lyophilized the CRISPR reagents in the reaction chamber and pre-stored the liquid solutions in blisters. We employed a low-cost, portable hand warmer to incubate the microfluidic chip without the need for electricity. The self-contained microfluidic system can detect down to 100 copies of SARS-CoV-2 RNA. Further, we clinically validated our method by detecting 24 COVID-19 clinical nasopharyngeal swab samples, achieving excellent sensitivity (94.1%), specificity (100%), and accuracy (95.8%). This simple, sensitive, and affordable microfluidic system represents a promising tool for point-of-care diagnostics of COVID-19 and other infectious diseases.
Keywords: CRISPR-Cas12a; Hand warmer; Instrument-free molecular diagnostics; SARS-CoV-2 detection; Self-contained microfluidic chip.
. 2021 Dec 8;199:113865.
doi: 10.1016/j.bios.2021.113865. Online ahead of print.
Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system
Ziyue Li[SUP] 1 [/SUP], Xiong Ding[SUP] 2 [/SUP], Kun Yin[SUP] 2 [/SUP], Lori Avery[SUP] 3 [/SUP], Enrique Ballesteros[SUP] 3 [/SUP], Changchun Liu[SUP] 4 [/SUP]
Affiliations
- PMID: 34906838
- DOI: 10.1016/j.bios.2021.113865
Abstract
Rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for early diagnostics and timely medical treatment of coronavirus disease 2019 (COVID-19). However, current detection methods typically rely on expensive and bulky instrumentation. Here, we developed a simple, sensitive, instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using a self-contained microfluidic system. The microfluidic chip integrates isothermal amplification, CRISPR cleavage, and lateral flow detection in a single, closed microfluidic platform, enabling contamination-free, visual detection. To simplify the operation and transportation of the device, we lyophilized the CRISPR reagents in the reaction chamber and pre-stored the liquid solutions in blisters. We employed a low-cost, portable hand warmer to incubate the microfluidic chip without the need for electricity. The self-contained microfluidic system can detect down to 100 copies of SARS-CoV-2 RNA. Further, we clinically validated our method by detecting 24 COVID-19 clinical nasopharyngeal swab samples, achieving excellent sensitivity (94.1%), specificity (100%), and accuracy (95.8%). This simple, sensitive, and affordable microfluidic system represents a promising tool for point-of-care diagnostics of COVID-19 and other infectious diseases.
Keywords: CRISPR-Cas12a; Hand warmer; Instrument-free molecular diagnostics; SARS-CoV-2 detection; Self-contained microfluidic chip.