Biosensors & bioelectronicsMar 7, 2025

Combined Rapid DNA Test for Detecting Monkeypox Virus in One Step

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Feb 16, 2026

Abstract

A CRISPR-Cas12a-based detection system for monkeypox virus can achieve a detection limit as low as 10 aM.

The system specifically targets the MPXV A27L gene and shows no cross-reactivity with other orthopoxviruses.
Results can be obtained in under 40 minutes, making it suitable for rapid testing.
Validation using six different clinical sample types indicates that herpes fluid and saliva are the most effective for detection.
Findings align with results from qPCR, suggesting comparable reliability.
Lyophilization of RPA and CRISPR reagents enhances transport, storage, and deployment capabilities.

Simplified

Monkeypox is a zoonotic disease caused by the monkeypox virus (MPXV), with outbreaks primarily occurring in West and Central Africa. The recent global MPXV outbreak underscores the urgent need for effective detection methods. Currently, qPCR is considered the gold standard for MPXV detection; however, it requires specialized personnel and costly equipment. This study introduces a CRISPR-Cas12a-based detection system targeting the MPXV A27L gene, achieving a detection limit as low as 10 aM. This system exhibits high specificity, with no cross-reactivity with other orthopoxviruses, and delivers results in under 40 min. To support point-of-care testing (POCT), we developed a lateral flow assay (LFA) strip for easy result visualization. The detection system was validated using six different clinical sample types, revealing that herpes fluid and saliva are the most suitable sources. The findings of this study align with qPCR results. Additionally, we lyophilized the RPA and CRISPR reagents to improve transport, storage, and field deployment. In conclusion, this study presents a reliable molecular diagnostic approach for early MPXV detection and point-of-care testing, contributing to epidemic prevention and control.

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Competing interests

Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

PubMed

Combined Rapid DNA Test for Detecting Monkeypox Virus in One Step

Abstract

Full Text

Funding

Competing interests

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