Multiple gRNAs-assisted CRISPR/Cas12a-based portable aptasensor enabling glucometer readout for amplification-free and quantitative detection of malathion

Yi Tian ¹, Jiaxuan Chen ¹, Fengzheng Chen ¹

⁰College of New Energy Materials and Chemistry, Leshan Normal University, Leshan, Sichuan, 614000, PR China; Sichuan Province Key Laboratory of Natural Products and Small Molecule Synthesis, Leshan, Sichuan, 614000, PR China.

Analytica Chimica Acta +

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January 29, 2025

View abstract on PubMed

Summary

A new multiplex-guide RNA-assisted CRISPR/Cas12a portable aptasensor (MgCPA) enables sensitive, amplification-free detection of malathion pesticide residues. This innovative method offers a promising tool for ensuring food safety by detecting malathion in various food samples.

Area Of Science

Biotechnology
Food Safety
Biosensor Technology

Background

Malathion pesticide residues pose significant food safety risks.
CRISPR/Cas systems offer innovative detection but haven't been applied to malathion.
Existing multiple-guide RNA CRISPR/Cas biosensors are limited to nucleic acid targets.

Purpose Of The Study

To develop a novel biosensor for the sensitive and quantitative detection of malathion.
To adapt CRISPR/Cas technology for non-nucleic acid targets like pesticides.
To create an amplification-free, portable detection method for malathion residues.

Main Methods

Development of a multiplex-guide RNA-assisted CRISPR/Cas12a portable aptasensor (MgCPA).
Utilized aptamer binding to activate CRISPR/Cas12a trans-cleavage activity in the presence of malathion.
Employed a glucometer for signal detection via cleavage of invertase-HP probes.

Main Results

Achieved amplification-free, quantitative detection of malathion down to 300 fM.
Demonstrated satisfactory selectivity, reproducibility, and stability of the MgCPA strategy.
Successfully applied the MgCPA strategy for malathion detection in real food samples (orange, apple, cabbage, spinach).

Conclusions

The MgCPA strategy provides a sensitive, portable, and selective method for malathion detection in food.
This work expands the application of multiple-guide RNA CRISPR/Cas12a systems to non-nucleic acid targets.
The developed aptasensor holds significant potential for enhancing food safety monitoring.

Keywords:

Amplification-free CRISPR/Cas12a Malathion Multiple gRNAs Portable quantitative detection