CRISPR/Cas12a dual-mode biosensor for Staphylococcus aureus detection via enzyme-free isothermal amplification

Hongmin Gao; Hehua Zhang; Xue Qi; Meng Miao; Longbin Que; Xin Gu; Dong Chang; Hongzhi Pan

doi:10.1016/j.talanta.2024.127013

CRISPR/Cas12a dual-mode biosensor for Staphylococcus aureus detection via enzyme-free isothermal amplification

Talanta. 2024 Oct 10:282:127013. doi: 10.1016/j.talanta.2024.127013. Online ahead of print.

Authors

Hongmin Gao¹, Hehua Zhang², Xue Qi², Meng Miao², Longbin Que¹, Xin Gu¹, Dong Chang³, Hongzhi Pan⁴

Affiliations

¹ School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai, 200093, People's Republic of China; Collaborative Research Center, Shanghai University of Medicine and Health Sciences, Shanghai, 201318, People's Republic of China.
² Collaborative Research Center, Shanghai University of Medicine and Health Sciences, Shanghai, 201318, People's Republic of China.
³ Department of Clinical Laboratory, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai, 201399, People's Republic of China. Electronic address: changdongydy@163.com.
⁴ Collaborative Research Center, Shanghai University of Medicine and Health Sciences, Shanghai, 201318, People's Republic of China; The Affiliated Zhoupu Hospital, Shanghai University of Medicine & Health Sciences, Shanghai, 201318, People's Republic of China. Electronic address: panhongzhilaoshi@163.com.

PMID: 39406093
DOI: 10.1016/j.talanta.2024.127013

Abstract

Accurate and reliable detection of Staphylococcus aureus (S. aureus) is essential for preventing infections, particularly in healthcare and food safety contexts. This work presents a novel dual-mode biosensor that integrates the CRISPR/Cas12a system with an enzyme-free isothermal amplification method for detecting S. aureus. Hybridization chain reaction (HCR) and catalytic hairpin assembly (CHA) amplify the aptamer-triggered response, significantly enhancing sensitivity. CRISPR/Cas12a's nuclease activity is utilized in two modes: cis cleavage generates a fluorescence signal, while trans cleavage produces an electrochemical signal, enabling dual-mode detection. The biosensor demonstrates outstanding performance, with a limit of detection (LOD) as low as 5.7 CFU mL^-1 in electrochemical mode and 133.7 CFU mL^-1 in fluorescence mode, showcasing excellent accuracy, stability, and sensitivity. It has been successfully applied to detecting actual samples, confirming its practical applicability. This innovative approach offers a powerful tool for the swift and precise identification of S. aureus and paves the way for developing next-generation dual-mode biosensors for various analytes. Future research will aim to simplify the detection process further, making it more accessible for use in resource-limited settings.

Keywords: CHA; CRISPR/Cas12a; Dual-mode biosensor; HCR; S. aureus.