Citation: Xinrui Zhang, Hui Chen, Tingting Fan, Xueming Zhu, Ying Tan, Dan Gao. A microfluidic biosensor based on multiple signal amplification for rapid and sensitive detection of E. coli O157:H7 and Staphylococcus aureus[J]. Chinese Chemical Letters, ;2026, 37(4): 111705. doi: 10.1016/j.cclet.2025.111705 shu

A microfluidic biosensor based on multiple signal amplification for rapid and sensitive detection of E. coli O157:H7 and Staphylococcus aureus

Xinrui Zhang ^{a, b} ,
Hui Chen ^a ,
Tingting Fan ^a ,
Xueming Zhu ^{a, b} ,
Ying Tan ^{a, *,} ,
Dan Gao ^{a, b, *,}

a.
The State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Biology, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, China
b.
Key Laboratory of Metabolomics at Shenzhen, Shenzhen 518055, China

tan.ying@sz.tsinghua.edu.cn

gao.dan@sz.tsinghua.edu.cn

Received Date: 6 March 2025
Revised Date: 30 July 2025
Accepted Date: 10 August 2025
Available Online: 11 August 2025

Figures(5)

E. coli O157:H7 and Staphylococcus aureus have emerged as significant foodborne pathogens, characterized by considerable incidence rates and mortality. Despite advancements, current detection methods are hindered by challenges in enhancing specificity and sensitivity. Herein, we introduced a cutting-edge biosensor that employs a novel CHA-coupled CRISPR multi-stage signal amplification technique for the rapid and ultra-sensitive detection of these two pathogens. This microfluidic device consisted of an upstream serpentine mixing channel and a downstream boat-shaped microcavity equipped with a micro-column array, facilitating efficient reagent mixing, robust CHA amplification, and CRISPR reactions. Multiple signal amplification was achieved through bacterial competitive binding triggered by catalytic hairpin assembly (CHA) and crRNA-mediated CRISPR reactions. Based on this platform, the detection of target bacteria is transformed into nucleic acid detection, with a maximum detection range of 134 CFU/mL for E. coli O157:H7 and 181 CFU/mL for Staphylococcus aureus, which were better or comparable to previously reported biosensors. The entire assay was completed within approximately 1.5 h, with a minimal sample volume requirement of just 10 µL. The biosensor exhibited a high recovery rate, ranging from 95% to 115%, and demonstrated excellent specificity towards the target bacteria. In summary, this biosensor offers a rapid, accurate, and highly sensitive tool for food safety and clinical diagnostics.
- Microfluidic chip,
- Catalytic hairpin assembly,
- CRISPR,
- Staphylococcus aureus,
- E. coli O157:H7
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