TalantaDec 30, 2025

Sensitive fluorescent detection of mycotoxin using a CRISPR-based copper nanocluster sensor without markers

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jan 3, 2026

Abstract

The biosensing platform achieved a limit of detection of 47.51 pg/mL for aflatoxin B1.

A CRISPR/Cas12a-mediated system was developed for detecting aflatoxin B1.
Copper nanoclusters were synthesized using marker-free DNA within 5 minutes, demonstrating strong fluorescence.
The presence of aflatoxin B1 initiated reactions that produced a detectable decrease in fluorescence.
High specificity for aflatoxin B1 was observed within a linear detection range of 0.05-10 ng/mL.
The platform offers a novel approach for detecting non-nucleic acid targets.

Simplified

The problem of mycotoxin contamination in foodstuffs has attracted widespread attention and posed a great threat to human health. Therefore, the sensitive and effective detection of mycotoxins is of great importance for preserving public health worldwide. In this study, a CRISPR/Cas12a-mediated marker-free fluorescent biosensing platform was constructed for highly sensitive and fast detection of aflatoxin B1. The copper nanoclusters were synthesized with marker-free DNA single strands within 5 min, showing the outstanding fluorescence properties. With the existence of aflatoxin B1, the released complementary DNA (cDNA) triggered multiple isothermal amplification reaction. Subsequently, the obtained amplification products triggered the trans-cleavage activity of CRISPR/Cas12a system, which degraded the DNA single strands for synthesis of copper nanoclusters, leading to a decreased fluorescent signal. Benefiting from good fluorescence properties of copper nanoclusters, the established biosensing platform for aflatoxin B1 detection had a high specificity and a limit of detection of 47.51 pg/mL in the linear range of 0.05-10 ng/mL. The proposed platform provided a new insight for the detection of non-nucleic acid targets.

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Sensitive fluorescent detection of mycotoxin using a CRISPR-based copper nanocluster sensor without markers

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