Rxivist logo

Dynamic aqueous multiphase reaction system for simple, sensitive and quantitative one-pot CRISPR-Cas12a based molecular diagnosis

By Kun Yin, Xiong Ding, Ziyue Li, Hui Zhao, Kumarasen Cooper, Changchun Liu

Posted 24 Mar 2020
bioRxiv DOI: 10.1101/2020.03.21.001644

Recently, CRISPR-Cas technology has opened a new era of nucleic acid-based molecular diagnostics. However, current CRISPR-Cas-based nucleic acid biosensing has largely a lack of the quantitative detection ability and typically requires separate manual operations. Herein, we reported a dynamic aqueous multiphase reaction (DAMR) system for simple, sensitive and quantitative one-pot CRISPR-Cas12a based molecular diagnosis by taking advantage of density difference of sucrose concentration. In the DAMR system, recombinase polymerase amplification (RPA) and CRISPR-Cas12a derived fluorescent detection occurred in spatially separated but connected aqueous phases. Our DAMR system was utilized to quantitatively detect human papillomavirus (HPV) 16 and 18 DNAs with sensitivities of 10 and 100 copies within less than one hour. Multiplex detection of HPV16/18 in clinical human swab samples were successfully achieved in the DAMR system using 3D-printed microfluidic device. Furthermore, we demonstrated that target DNA in real human plasma samples can be directly amplified and detected in the DAMR system without complicated sample pre-treatment. As demonstrated, the DAMR system has shown great potential for development of next-generation point-of-care molecular diagnostics.

Download data

  • Downloaded 552 times
  • Download rankings, all-time:
    • Site-wide: 59,665
    • In bioengineering: 1,144
  • Year to date:
    • Site-wide: 55,244
  • Since beginning of last month:
    • Site-wide: 56,652

Altmetric data


Downloads over time

Distribution of downloads per paper, site-wide


PanLingua

News