Development and optimisation of a rapid and sensitive detection platform for Escherichia coli O157:H7 using RPA- Pf Ago
Year: 2025
Type: article
Abstract: Escherichia coli O157:H7 (E. coli O157:H7) is a significant foodborne pathogen that causes severe illnesses worldwide. In this study, a novel detection system combining recombinase polymerase amplification and Pyrococcus furiosus Argonaute (PfAgo) was developed for the rapid and sensitive identification of the conserved rfbE gene. The system integrates dual-fluorescence probes labelled with 6-carboxyfluorescein (FAM) and X-rhodamine (ROX) for parallel signal verification, which enhances detection reliability and minimises the risk of false positives. The optimised system achieved a limit of detection of 6.14 × 101 copies/μl, 100% specificity and repeatability, with a coefficient of variation below 10%. Validation using artificially contaminated milk and bottled water samples demonstrated complete agreement with the results of polymerase chain reaction, yielding 100% accuracy. With high sensitivity and specificity, this system enables rapid, accurate point-of-care screening for food safety and clinical diagnostics, offering broad potential in public health surveillance and foodborne pathogen control.
Authors: Yaqun Liu, Lianghui Chen, Zhiruo Wan, Jinkun Han, Xin Cai, Yongping Huang, Peikui Yang, Yicun Chen, Chengsong Xie, Zhenxia Zhang, Yuzhong Zheng
Language: English
Subfield: Biomedical Engineering
Field: Engineering
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $2,638
Funder: Hanshan Normal University
OpenAlex ID: W7119507995