A competitive aptamer binding-based CRISPR-cas biosensor for sensitive detection of tetracycline residues in biological samples

Yali Zhang; Xingpeng Dong; Cuijuan Jiang; Yanyan Yu; Haiyan Zhang; Jianjie Fu; Gaoxing Su; Yin Liu

doi:10.1016/j.talanta.2024.127491

A competitive aptamer binding-based CRISPR-cas biosensor for sensitive detection of tetracycline residues in biological samples

Talanta. 2024 Dec 29:286:127491. doi: 10.1016/j.talanta.2024.127491. Online ahead of print.

Authors

Yali Zhang¹, Xingpeng Dong², Cuijuan Jiang³, Yanyan Yu¹, Haiyan Zhang⁴, Jianjie Fu⁵, Gaoxing Su⁶, Yin Liu⁷

Affiliations

¹ Key Laboratory of Inflammation and Molecular Drug Targets of Jiangsu Province, School of Medicine, School of Pharmacy, Nantong University, Nantong 226001, China.
² School of Environment, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou 310024, China; Key Laboratory of Inflammation and Molecular Drug Targets of Jiangsu Province, School of Medicine, School of Pharmacy, Nantong University, Nantong 226001, China.
³ School of Environmental Science and Engineering, Shandong University, Qingdao 266237, China.
⁴ School of Environment, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou 310024, China.
⁵ School of Environment, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou 310024, China; State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
⁶ School of Environment, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou 310024, China; Key Laboratory of Inflammation and Molecular Drug Targets of Jiangsu Province, School of Medicine, School of Pharmacy, Nantong University, Nantong 226001, China. Electronic address: [email protected].
⁷ School of Environment, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou 310024, China. Electronic address: [email protected].

PMID: 39753078
DOI: 10.1016/j.talanta.2024.127491

Abstract

Tetracycline (TC) is widely used in veterinary medicine and animal feed; however, TC residues in food pose a risk to human health. Thus, the sensitive and selective detection of TC is needed to ensure food safety. Herein, we developed a CRISPR-Cas12a biosensor with competitive aptamer binding to detect TC residues. The aptasensor, formed by hybridizing activator DNA with TC-specific aptamers on streptavidin-modified magnetic beads, releases activator DNA in a TC concentration-dependent manner. This activated the Cas12a-crRNA complex, which cleaved single-strand DNA reporters to generate a detectable fluorescence signal. The TC signal was amplified through a two-step incubation reaction, with a detection limit as low as 9.45 × 10^-5 μg L^-1. The assay showed high selectivity and good recovery rates in various biological samples (e.g., honey, milk, fish), demonstrating the applicability of the biosensors in pollutant detection.

Keywords: Aptasensor; Biological sample; CRISPR/Cas12a; Pollutant detection; Tetracycline.