Amplification-free detection of mycoplasma pneumoniae via CRISPR-Cas12a and deep learning-optimized crRNAs on a lateral flow platform

Oct 23, 2025Journal of pharmaceutical and biomedical analysis

Direct detection of Mycoplasma pneumoniae using CRISPR-Cas12a and AI-optimized guides on a paper strip test

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Abstract

The multiplexed crRNA library improved sensitivity by 16.8-fold, achieving a limit of detection of 0.15 pM.

  • An amplification-free CRISPR-Cas12a detection platform was developed for Mycoplasma pneumoniae infection.
  • The system uses a deep learning-optimized crRNA library targeting conserved regions of the MP P1 gene.
  • Combinatorial use of 16 highly active crRNAs enhanced fluorescence signal intensity and reaction kinetics.
  • The assay demonstrated a tenfold increase in visual detection sensitivity when deployed on lateral flow strips.
  • Clinical evaluations showed over 95% agreement with standard clinical results and high specificity, with no cross-reactivity to other pathogens.

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