CRISPR/Cas12a and G-quadruplex DNAzyme-driven multimodal biosensor for visual detection of Aflatoxin B1

Zhihui Wu; Da-Wen Sun; Hongbin Pu

doi:10.1016/j.saa.2023.123121

CRISPR/Cas12a and G-quadruplex DNAzyme-driven multimodal biosensor for visual detection of Aflatoxin B1

Spectrochim Acta A Mol Biomol Spectrosc. 2023 Dec 5:302:123121. doi: 10.1016/j.saa.2023.123121. Epub 2023 Jul 8.

Authors

Zhihui Wu¹, Da-Wen Sun², Hongbin Pu¹

Affiliations

¹ School of Food Science and Engineering, South China University of Technology, Guangzhou 510641, China; Academy of Contemporary Food Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Guangzhou 510006, China; Engineering and Technological Research Centre of Guangdong Province on Intelligent Sensing and Process Control of Cold Chain Foods, & Guangdong Province Engineering Laboratory for Intelligent Cold Chain Logistics Equipment for Agricultural Products, Guangzhou Higher Education Mega Centre, Guangzhou 510006, China.
² School of Food Science and Engineering, South China University of Technology, Guangzhou 510641, China; Academy of Contemporary Food Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Guangzhou 510006, China; Engineering and Technological Research Centre of Guangdong Province on Intelligent Sensing and Process Control of Cold Chain Foods, & Guangdong Province Engineering Laboratory for Intelligent Cold Chain Logistics Equipment for Agricultural Products, Guangzhou Higher Education Mega Centre, Guangzhou 510006, China; Food Refrigeration and Computerized Food Technology (FRCFT), Agriculture and Food Science Centre, University College Dublin, National University of Ireland, Belfield, Dublin 4, Ireland. Electronic address: dawen.sun@ucd.ie.

PMID: 37579713
DOI: 10.1016/j.saa.2023.123121

Abstract

Aflatoxin B1 (AFB1) contamination severely threatens human and animal health, it is thus critical to construct a strategy for its rapid, accurate, and visual detection. Herein, a multimodal biosensor was proposed based on CRISPR/Cas12a cleaved G-quadruplex (G4) for AFB1 detection. Briefly, specific binding of AFB1 to the aptamer occupied the binding site of the complementary DNA (cDNA), and cDNA then activated Cas12a to cleave G4 into fragments. Meanwhile, the intact G4-DNAzyme could catalyze 3, 3', 5, 5'-tetramethylbenzidine (TMB) to form colourimetric/SERS/fluorescent signal-enhanced TMBox, and the yellow solution produced by TMBox under acidic conditions could be integrated with a smartphone application for visual detection. The colourimetric/SERS/fluorescent biosensor yielded detection limits of 0.85, 0.79, and 1.65 pg·mL^-1, respectively, and was applied for detecting AFB1 in peanut, maize, and badam samples. The method is suitable for visual detection in naturally contaminated peanut samples and has prospective applications in the food industry.

Keywords: Aflatoxin B1; Biosensor; CRISPR/Cas12a; G4-DNAzyme; Multimodal.

MeSH terms

Aflatoxin B1 / analysis
Aptamers, Nucleotide* / chemistry
Biosensing Techniques* / methods
CRISPR-Cas Systems
Coloring Agents
DNA, Catalytic*
DNA, Complementary / chemistry
DNA, Complementary / genetics
Food Contamination / analysis
Humans
Limit of Detection

Substances

Aflatoxin B1
DNA, Complementary
DNA, Catalytic
Aptamers, Nucleotide
Coloring Agents