4.8 Article

Enhanced Isothermal Amplification for Ultrafast Sensing of SARS-CoV-2 in Microdroplets

期刊

ANALYTICAL CHEMISTRY
卷 94, 期 10, 页码 4135-4140

出版社

AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.2c00008

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资金

  1. China Postdoctoral Science Foundation [2021TQ0210]
  2. National Natural Science Foundation of China [21804007, 21890742]
  3. SZU Top Ranking Project [86000000210]
  4. Shenzhen Stability Support Plan [20200806163622001]
  5. Shenzhen Key Laboratory for NanoBiosensing Technology [ZDSYS20210112161400001]
  6. Shenzhen Overseas Talent Program

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This study reports an integrated microdroplet array platform composed of an ultrasonic unit and minipillar array for enhanced RPA detection of SARS-CoV-2. The independent microvolume reactions and microstreaming driven by ultrasound greatly accelerate the amplification and increase the sensitivity, while reducing reagent consumption.
Rapid and high-throughput screening is critical to control the COVID-19 pandemic. Recombinase polymerase amplification (RPA) with highly accessible and sensitive nucleic acid amplification has been widely used for point-of-care infection diagnosis. Here, we report an integrated microdroplet array platform composed of an ultrasonic unit and minipillar array to enhance the RPA for ultrafast, high-sensitivity, and high-throughput detection of SARS-CoV-2. On such a platform, the independent microvolume reactions on individual minipillars greatly decrease the consumption of reagents. The microstreaming driven by ultrasound creates on-demand contactless microagitation in the microdroplets and promotes the interaction between RPA components, thus greatly accelerating the amplification. In the presence of microstreaming, the detection time is 6-12 min, which is 38.8-59.3% shorter than that of controls without microstreaming, and the end-point fluorescence intensity also increased 1.3-1.7 times. Furthermore, the microagitation-enhanced RPA also exhibits a lower detection limit (0.42 copy/mu L) for SARS-CoV-2 in comparison to the controls. This integrated microdroplet array detection platform is expected to meet the needs for high-throughput nucleic acid testing (NAT) to improve the containment of viral transmission during the epidemic, as well as provide a potential platform for the timely detection of other pathogens or viruses.

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