4.6 Article

Multiplex Digital MicroRNA Detection Using Cross-Inhibitory DNA Circuits

期刊

ACS SENSORS
卷 5, 期 8, 页码 2430-2437

出版社

AMER CHEMICAL SOC
DOI: 10.1021/acssensors.0c00593

关键词

multiplex assay; DNA circuit; microRNA; digital droplet detection; isothermal amplification

资金

  1. Universite de Recherche Paris Sciences et Lettres (PSL, prematuration program Digiplex)
  2. ESPCI-Paris
  3. Centre National de la Recherche Scientifique (CNRS)
  4. European Research Council [ProFF 647275, Deepmir 780519]

向作者/读者索取更多资源

Ubiquitous post-transcriptional regulators in eukaryotes, microRNAs are currently emerging as promising biomarkers of physiological and pathological processes. Multiplex and digital detection of microRNAs represents a major challenge toward the use of microRNA signatures in clinical settings. The classical reverse transcription polymerase chain reaction quantification approach has important limitations because of the need for thermocycling and a reverse transcription step. Simpler, isothermal alternatives have been proposed, yet none could be adapted in both a digital and multiplex format. This is either because of a lack of sensitivity that forbids single molecule detection or molecular cross-talk reactions that are responsible for nonspecific amplification. Building on an ultrasensitive isothermal amplification mechanism, we present a strategy to suppress cross-talk reactions, allowing for robust isothermal and multiplex detection of microRNA targets. Our approach relies on target-specific DNA circuits interconnected with DNA-encoded inhibitors that repress nonspecific signal amplification. We demonstrate the one-step, isothermal, digital, and simultaneous quantification of various pairs of important microRNA targets.

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