Microfluidic space coding for multiplexed nucleic acid detection via CRISPR-Cas12a and recombinase polymerase amplification

Zhichen Xu, Dongjuan Chen, Tao Li, Jiayu Yan, Jiang Zhu, Ting He, Rui Hu, Ying Li (), Yunhuang Yang and Maili Liu
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Zhichen Xu: Chinese Academy of Sciences
Dongjuan Chen: Huazhong University of Science and Technology
Tao Li: Chinese Academy of Sciences
Jiayu Yan: Chinese Academy of Sciences
Jiang Zhu: Chinese Academy of Sciences
Ting He: Chinese Academy of Sciences
Rui Hu: Chinese Academy of Sciences
Ying Li: Chinese Academy of Sciences
Yunhuang Yang: Chinese Academy of Sciences
Maili Liu: Chinese Academy of Sciences

Nature Communications, 2022, vol. 13, issue 1, 1-14

Abstract: Abstract Fast, inexpensive, and multiplexed detection of multiple nucleic acids is of great importance to human health, yet it still represents a significant challenge. Herein, we propose a nucleic acid testing platform, named MiCaR, which couples a microfluidic device with CRISPR-Cas12a and multiplex recombinase polymerase amplification. With only one fluorescence probe, MiCaR can simultaneously test up to 30 nucleic acid targets through microfluidic space coding. The detection limit achieves 0.26 attomole, and the multiplexed assay takes only 40 min. We demonstrate the utility of MiCaR by efficiently detecting the nine HPV subtypes targeted by the 9-valent HPV vaccine, showing a sensitivity of 97.8% and specificity of 98.1% in the testing of 100 patient samples at risk for HPV infection. Additionally, we also show the generalizability of our approach by successfully testing eight of the most clinically relevant respiratory viruses. We anticipate this effective, undecorated and versatile platform to be widely used in multiplexed nucleic acid detection.

Date: 2022
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DOI: 10.1038/s41467-022-34086-y

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