Method for detecting mycotoxins based on the crisper-cas12a system

The electrochemical sensor based on the CRISPR-Cas12a system utilizes the reaction between single-stranded DNA and mycotoxin recognition DNA to form a complex, which combines with helper DNA and enzymatically activated DNA. This solves the problems of insufficient sensitivity and complex operation in the detection of mycotoxins in food, achieving high sensitivity and high selectivity in detection.

CN117470930BActive Publication Date: 2026-06-16ZHEJIANG LAB +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
ZHEJIANG LAB
Filing Date
2022-07-27
Publication Date
2026-06-16

AI Technical Summary

⚠Technical Problem

Existing technologies for detecting mycotoxins in food lack sufficient sensitivity and are complex to operate, making them unsuitable for detecting complex food matrices.

⚗Method used

An electrochemical sensor based on the CRISPR-Cas12a system was used to detect fungal toxins by reacting single-stranded DNA with fungal toxin recognition DNA to form a complex, which then binds to helper DNA and is activated by enzymatic digestion. The trans-cleavage activity of CRISPR-Cas12a was utilized to detect fungal toxins in the electrochemical biosensor.

🎯Benefits of technology

It achieves highly sensitive and selective detection of mycotoxins, is simple to operate, is suitable for complex food matrices, has a detection range of 1×10-6-5 ng/mL, a detection limit of 0.74 fg/mL, and is applicable to the detection of a variety of mycotoxins.

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Abstract

The present application relates to the field of electrochemical analysis and detection, and discloses a method for detecting mycotoxin based on CRISPR-Cas12a system, comprising: (1) reacting single-stranded DNA with recognition DNA of mycotoxin to obtain single-stranded-recognition DNA complex, wherein the single-stranded DNA is complementary to the recognition DNA; (2) mixing the single-stranded-recognition DNA complex and the sample to be tested which may contain mycotoxin to obtain reaction mixture I after reaction II, and mixing the reaction mixture I with auxiliary DNA containing activated DNA fragments to obtain reaction mixture II after reaction III, wherein the auxiliary DNA is complementary to the single-stranded DNA; (3) mixing the reaction mixture II with CRISPR-Cas12a system solution to obtain reaction solution after activation; (4) adding the reaction solution dropwise to an electrochemical biosensor to perform reaction V, and detecting the electrochemical signal of the electrochemical biosensor after reaction V is completed. The method has high sensitivity and high selectivity, and is simple and rapid to operate.
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