Related Experiment Video
Updated: Jun 5, 2025
04:33
Author Spotlight: Engineering Molecular Tools for Disease Detection and Imaging
Published on: December 8, 2023
764
Novel Multimode Assay Based on Asymmetrically Competitive CRISPR and Raman Barcode Spectra for Multiple
Hao Jiang1, Cheng Qian1, Yulin Deng1
1Beijing Key Laboratory for Separation and Analysis in Biomedicine and Pharmaceuticals, School of Medical Technology, Beijing Institute of Technology, Beijing 100081, China.
Analytical Chemistry
|December 6, 2024
Summary
This study introduces a novel assay combining asymmetric competitive CRISPR (acCRISPR) and surface-enhanced Raman spectroscopy with pregnancy test strips (PTS) for sensitive, multiplexed detection of hepatocellular carcinoma biomarkers.
Area of Science:
- Biomarker detection
- Biosensing technologies
- Molecular diagnostics
Background:
- Commercial pregnancy test strips (PTS) offer advantages like cost-effectiveness and stability, making them attractive platforms for novel sensing strategies.
- Current PTS integrations face limitations in quantification, sensitivity, and multiplexing capabilities for disease biomarker detection.
- Hepatocellular carcinoma (HCC) biomarker detection requires sensitive and specific methods for early diagnosis.
Purpose of the Study:
- To develop a sensitive and multiplexed assay for detecting hepatocellular carcinoma (HCC) biomarkers miRNA122 and miRNA233.
- To overcome the limitations of traditional PTS by enhancing quantification, sensitivity, and multiple detection capabilities.
- To integrate asymmetric competitive CRISPR (acCRISPR) and surface-enhanced Raman spectroscopy (SERS) with PTS for a novel diagnostic approach.
Main Methods:
- Developed the asymmetric competitive CRISPR (acCRISPR) assay as a non-amplified, cascaded signal amplification method.
- Utilized two distinct AuNPs-based core-shell Raman tags for visual recognition and spectral segmentation of biomarkers.
- Coupled acCRISPR and SERS with PTS, creating the acCRISPR-PTS-SERS assay for dual-signal detection.
- Optimized assay conditions to achieve high sensitivity and specificity for miRNA detection.
Main Results:
- Achieved low limits of detection (LOD) for miRNA122 (10.36 fM) and miRNA233 (4.65 fM).
- Demonstrated a sensitivity enhancement of nearly two orders of magnitude compared to existing methods.
- Successfully enabled visual classification recognition and spectral signal subdivision for multiplexed biomarker detection.
- Validated the assay's capability for enhanced quantification and multiple detection using dual signals.
Conclusions:
- The acCRISPR-PTS-SERS assay offers a highly sensitive and multiplexed approach for detecting HCC biomarkers.
- This novel integration overcomes key limitations of current PTS-based sensing strategies.
- The developed assay holds potential for future development of hand-held, smartphone-assisted diagnostic tools for various cancers.
