Work

Microfluidic Padlock Probe-based Rolling Circle Amplification for sensitive detection of mecA resistance gene in Staphylococcus aureus

Year: 2025

Type: article

Abstract: Antimicrobial resistance and the dissemination of resistance genes remain critical global health challenges, demanding efficient monitoring and detection tools. Methicillin-resistant Staphylococcus aureus (MRSA) is a prominent example, posing serious clinical and epidemiological concerns. Here, we developed a sensitive detection platform combining isothermal Padlock Probe-based Rolling Circle Amplification with a bead-based microfluidic biosensor and fluorescent readout for the detection of the mecA gene in S. aureus . The assay design was first validated using a complementary synthetic ssDNA target, then optimized with genomic dsDNA extracted from a laboratory strain. The optimized system was subsequently applied to clinical isolates from sepsis cases and complex matrices simulating natural habitats, including milk, serum, and salad. The biosensor achieved a detection limit of 10 4 cells with 100 % specificity compared to PCR at an analysis time of two hour. All reaction steps were carried out sequentially within the microfluidic device, highlighting the strong potential of the device for full automation and for extension to additional bacterial genes.

Source: Sensing and Bio-Sensing Research

DOI: 10.1016/j.sbsr.2025.100941

Authors: Léo Baldenweck, Catarina Caneira, Jasmina Vidic

Institutions: AgroParisTech, Université Paris-Saclay, Microbiologie de l’alimentation au service de la santé, Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement, Instituto de Engenharia de Sistemas e Computadores Microsistemas e Nanotecnologias

Language: English

Topic: Biosensors and Analytical Detection

Subfield: Biomedical Engineering

Field: Engineering

Domain: Physical Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $2,850