Two-Dimensional Nanozyme-Catalyzed Colorimetric CRISPR Assay for the Microfluidic Detection of Monkeypox Virus

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Diagnostics

Background:

• The global monkeypox virus (MPXV) outbreaks necessitate rapid and precise diagnostic methods, particularly in resource-limited environments.
• Existing diagnostic techniques may face challenges in speed, sensitivity, or accessibility in diverse settings.

Purpose of the Study:

• To develop a novel two-dimensional nanozyme-catalyzed colorimetric CRISPR assay for microfluidic detection of MPXV.
• To evaluate the assay's sensitivity, speed, and accuracy using clinical samples.

Main Methods:

• Preparation of two-dimensional GO@Pt nanomaterials via graphene oxide substrate, gold seed adsorption, and porous Pt shell deposition.
• A single-step recombinase polymerase amplification-CRISPR/Cas13a assay for MPXV nucleic acid detection.
• Colorimetric detection using GO@Pt nanozymes, FAM-biotin labeled ssRNA reporters, and TMB substrate.

Main Results:

• The GO@Pt-catalyzed colorimetric CRISPR assay achieved a limit of detection of 1 copy/microL of MPXV.
• The assay provided results within 60 minutes.
• Testing on forty clinical samples demonstrated 100% agreement with real-time PCR results.

Conclusions:

• The developed GO@Pt-catalyzed colorimetric CRISPR assay is a sensitive and accurate diagnostic tool for MPXV detection.
• The microfluidic and colorimetric nature of the assay makes it highly suitable for resource-constrained settings.
• This assay holds significant potential for improving MPXV surveillance and diagnosis globally.