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Updated: May 31, 2026

Point-of-care CRISPR-based Diagnostics with Premixed and Freeze-dried Reagents
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Ultrasensitive Wash-Free Homogeneous CRISPR Assay Using Spatial Proximity Chemiluminescence Reporter.

Yu Zhang1, Liangwen Hao2, Quanshuang Li3

  • 1Department of Medical Imaging, Tongji Hospital, Shanghai Frontiers Science Center of Nanocatalytic Medicine, The Institute for Biomedical Engineering & Nano Science, School of Medicine, Tongji University, Shanghai 200065, China.

ACS Nano
|May 29, 2026
PubMed
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This summary is machine-generated.

This study introduces a novel spatial proximity chemiluminescence (SPC) reporter for CRISPR-Cas12a diagnostics, significantly enhancing sensitivity for point-of-care testing. The SPC-CRISPR platform offers attomolar detection limits and simplified workflows for molecular diagnostics.

Area of Science:

  • Molecular Diagnostics
  • Biotechnology
  • Chemical Biology

Background:

  • CRISPR-based diagnostics offer potential for point-of-care (POC) testing but face challenges with sensitivity and workflow complexity.
  • Existing CRISPR diagnostic methods often require complex sample preparation and lack sufficient sensitivity for early disease detection.

Purpose of the Study:

  • To develop a highly sensitive and simplified CRISPR-based diagnostic platform for POC applications.
  • To create a novel reporter system that converts CRISPR-Cas12a activity into a robust, excitation-free chemiluminescent signal.
  • To validate the diagnostic performance of the new platform using clinical samples.

Main Methods:

  • Development of a spatial proximity chemiluminescence (SPC) reporter system for CRISPR-Cas12a.
Keywords:
CRISPRPOCmatrix interferencespatial proximity chemiluminescencestreamlined workflow

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  • Integration of SPC reporter with CRISPR-Cas12a for attomolar detection limits and enhanced sensitivity.
  • Combination with recombinase polymerase amplification for single-tube, highly sensitive assays.
  • Validation using clinical samples for HPV16 and cancer-associated microRNA detection.
  • Implementation in a portable, microfluidic-based device for sample-to-result diagnostics.

    • Main Results:

    • The SPC-CRISPR system achieved attomolar-level detection limits with a 50,000-fold sensitivity enhancement.
    • Demonstrated an 8-log dynamic range suitable for quantitative analysis with minimal sample preprocessing.
    • Achieved 99.2% accuracy for HPV16 detection in cervical swabs and 86.4% accuracy for miR-19a in plasma samples.
    • The portable device showed 100% concordance with quantitative PCR (qPCR) results.

    Conclusions:

    • The SPC-CRISPR platform significantly enhances sensitivity and simplifies workflows for molecular diagnostics.
    • This technology holds great potential for accurate and accessible point-of-care molecular testing.
    • The developed diagnostic device offers a wash-free, homogeneous, and portable solution for sample-to-result analysis.