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CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...

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Related Experiment Video

Updated: May 23, 2025

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

Published on: March 9, 2017

Origami Paper-Based Immunoassay Device with CRISPR/Cas12a Signal Amplification.

Hikaru Suzuki¹, Guodong Tong¹, Pabitra Nath²

¹Department of Applied Chemistry, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Kanagawa 223-8522, Japan.

|March 10, 2025

Summary

This summary is machine-generated.

This study integrates CRISPR/Cas12a with paper-based ELISA for rapid, sensitive protein detection. The new method significantly improves detection limits for targets like hepatitis B surface antigen, enabling point-of-care diagnostics.

Keywords:

CRISPR/Cas ELISA fluorescence immunoassay paper-based analytical devices

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Area of Science:

Biotechnology
Molecular Diagnostics
Assay Development

Background:

Conventional enzyme-linked immunosorbent assays (ELISA) are time-consuming for clinical diagnosis.
Paper-based ELISA offers faster, cheaper analysis but often lacks sensitivity.
Limited sensitivity in paper-based assays hinders clinical application for low-concentration targets.

Purpose of the Study:

To develop a simple, rapid, and highly sensitive detection method for non-nucleic acid targets.
To integrate the CRISPR/Cas12a system into a paper-based ELISA platform.
To enhance the sensitivity of paper-based immunoassays for clinical diagnosis.

Main Methods:

Designed an origami-type paper-based device for assay operation.
Utilized CRISPR/Cas12a enzyme with a fluorophore/quencher-labeled ssDNA probe for fluorescence detection.
Employed antibodies labeled with DNA to activate CRISPR/Cas12a for enhanced signal amplification.

Main Results:

Achieved significantly boosted detection sensitivity for human IgG and hepatitis B virus surface antigen (HBsAg).
Demonstrated a limit of detection (LOD) of 12 pg/mL for HBsAg, over 10-fold improvement compared to commercial ELISA kits (200 pg/mL).
Validated fluorescence detection in porcine whole blood samples using smartphone imaging for quantitative analysis.

Conclusions:

The developed CRISPR/Cas12a-integrated paper-based ELISA platform offers high sensitivity and rapid detection.
The platform shows potential for point-of-care clinical diagnostics, overcoming limitations of existing paper-based assays.
This approach facilitates sensitive detection of non-nucleic acid targets in complex biological samples.