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November 12, 2025Analytical Chemistry

Allele-Specific CRISPR-Cas9-Based Ratiometric Fluorescence Platform for Portable EGFR L858R Mutation Detection

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Authors

WMWenjun MingYZYidan ZhuLLLongjie Li

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Overview

This platform detects circulating tumor DNA mutations in point-of-care testing, indicating full clinical concordance with diagnoses.

Key Points

  • To develop a portable fluorescence platform for detecting low-abundance EGFR L858R mutations.
  • Utilized a CRISPR-Cas9-based ratiometric fluorescence sensor.
  • Engineered sgRNA to position the L858R mutation in the PAM region of Cas9.
  • Achieved visualization of mutation at 0.01% allele frequency using a 3D-printed device.
  • Demonstrated 100% concordance with clinical diagnoses based on 22 plasma samples.
  • Achieved single-nucleotide specificity in mutation detection.

Cite This Study

Ming et al. (2025) studied this question.

synapsesocial.com/papers/692523d9c0ce034ddc3558e4https://doi.org/10.1021/acs.analchem.5c05548
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