Au-Fe3O4 nanozyme coupled with CRISPR-Cas12a for sensitive and visual antibiotic resistance diagnosing

Haoxiang Chen; Bangying Li; Shangyi Shi; Tao Zhou; Xiumin Wang; Zuyong Wang; Xi Zhou; Miao Wang; Wei Shi; Lei Ren

doi:10.1016/j.aca.2023.341014

Au-Fe₃O₄ nanozyme coupled with CRISPR-Cas12a for sensitive and visual antibiotic resistance diagnosing

Anal Chim Acta. 2023 Apr 22:1251:341014. doi: 10.1016/j.aca.2023.341014. Epub 2023 Feb 25.

Authors

Haoxiang Chen¹, Bangying Li², Shangyi Shi³, Tao Zhou¹, Xiumin Wang², Zuyong Wang⁴, Xi Zhou¹, Miao Wang¹, Wei Shi⁵, Lei Ren⁶

Affiliations

¹ Department of Biomaterials, The Higher Educational Key Laboratory for Biomedical Engineering of Fujian Province, Research Center of Biomedical Engineering of Xiamen, College of Materials, Xiamen University, Xiamen, 361005, PR China.
² School of Pharmaceutical Sciences, Xiamen University, Xiamen, 361102, PR China.
³ State Key Laboratory of Computer Architecture, Institute of Computing Technology, Chinese Academy of Sciences, Beijing, 100190, PR China; School of Computer Science and Technology, University of Chinese Academy of Sciences, Beijing, 100049, PR China.
⁴ College of Materials Science and Engineering, Hunan University, Changsha, 410072, PR China.
⁵ Department of Biomaterials, The Higher Educational Key Laboratory for Biomedical Engineering of Fujian Province, Research Center of Biomedical Engineering of Xiamen, College of Materials, Xiamen University, Xiamen, 361005, PR China. Electronic address: shiwei@xmu.edu.cn.
⁶ Department of Biomaterials, The Higher Educational Key Laboratory for Biomedical Engineering of Fujian Province, Research Center of Biomedical Engineering of Xiamen, College of Materials, Xiamen University, Xiamen, 361005, PR China; State Key Lab of Physical Chemistry of Solid Surfaces, Xiamen University, Xiamen, 361005, PR China. Electronic address: renlei@xmu.edu.cn.

PMID: 36925313
DOI: 10.1016/j.aca.2023.341014

Abstract

The accumulation and spread of antibiotic resistance bacteria (ARB) in the environment may accelerate the formation of superbugs and seriously threaten the health of all living beings. The timeliness and accurate diagnosing of antibiotic resistance is essential to controlling the propagation of superbugs in the environment and formulating effective public health management programs. Herein, we developed a speedy, sensitive, accurate, and user-friendly colorimetric assay for antibiotic resistance, via a synergistic combination of the peroxidase-like property of the Au-Fe₃O₄ nanozyme and the specific gene identification capability of the CRISPR-Cas12a. Once the CRISPR-Cas12a system recognizes a target resistance gene, it activates its trans-cleavage activity and subsequently releases the Au-Fe₃O₄ nanozymes, which oxidizes the 3,3,5,5-tetramethylbenzidine (TMB) with color change from transparent to blue. The diagnosing signals could be captured and analyzed by a smartphone. This method detected kanamycin-resistance genes, ampicillin-resistance genes, and chloramphenicol-resistance genes by simple operation steps with high sensitivity (<0.1 CFU μL^-1) and speediness (<1 h). This approach may prove easy for the accurate and sensitive diagnosis of the ARGs or ARB in the field, thus surveilling and controlling the microbial water quality flexibly and efficiently.

Keywords: Antibiotic resistance bacteria; Antibiotic resistance gene; CRISPR/Cas12a; Colorimetric assay; Nanozyme; Pathogen detection.

MeSH terms

Ampicillin
Angiotensin Receptor Antagonists*
Angiotensin-Converting Enzyme Inhibitors*
CRISPR-Cas Systems
Drug Resistance, Microbial / genetics

Substances

Angiotensin Receptor Antagonists
Angiotensin-Converting Enzyme Inhibitors
Ampicillin