Combining tag-specific primer extension and magneto-DNA system for Cas14a-based universal bacterial diagnostic platform

作者：Song, Fengge; Wei, Yangdao; Wang, Peng; Ge, Xiaolin; Li, Chaoyang; Wang, Aimin; Yang, Zhiqing^*; Wan, Yi^*; Li, Jinghong

来源：Biosensors and Bioelectronics, 2021, 185: 113262.

DOI：10.1016/j.bios.2021.113262

• Bacterial diagnostics
• CRISPR-Cas14a
• Tag-specific primer extension
• Magneto-DNA

Nucleic acid-based diagnosis using CRISPR-Cas associated enzymes is essential for rapid infectious disease diagnosis and treatment strategies during a global pandemic. The obstacle has been blossomed CRIPSR-Cas based tools that can monitor wide range of pathogens in clinical samples with ultralow concentrations. Here, a universal nucleic acid magneto-DNA nanoparticle system was exploited for the detection of pathogenic bacteria, based on the collateral cleavage activity of CRISPR-Cas14a and tag-specific primer extension. In the system, the target nucleic acids were amplificated and be separated from mixtures by streptavidin-coated magnetic bead. The collateral cleavage activity of CRISPR-Cas14a can be activated via the tag sequence on the target product. Consequently, the fluorophore quencher reporter can be activated by CRISPR-Cas14a, leading to the increasing response. The exploited universal bacterial diagnostic can distinguish six different bacteria strains with 1 cfu/mL or 1 aM sensitivity, which may provide new strategies to construct fast, accurate, cost-effective and sensitive diagnostic tools in environments with limited resources.

• 单位
  中国科学院; y; 中国科学院海洋研究所; 海南大学; 清华大学