{"entity": "publication", "iuid": "590f8458b6a14cd0a3ffa29c82cde109", "timestamp": "2026-08-20T20:42:30.122Z", "links": {"self": {"href": "https://publications-affiliated.scilifelab.se/publication/590f8458b6a14cd0a3ffa29c82cde109.json"}, "display": {"href": "https://publications-affiliated.scilifelab.se/publication/590f8458b6a14cd0a3ffa29c82cde109"}}, "title": "A systematic implementation of padlock probing-based rolling circle amplification in an integrated microfluidic device for quantitative biomolecular analyses.", "authors": [{"family": "Caneira", "given": "Catarina R F", "initials": "CRF"}, {"family": "Rosa", "given": "Rafaela R", "initials": "RR"}, {"family": "Chu", "given": "Virginia", "initials": "V"}, {"family": "Nilsson", "given": "Mats", "initials": "M"}, {"family": "Madaboosi", "given": "Narayanan", "initials": "N"}, {"family": "Soares", "given": "Ruben R G", "initials": "RRG"}, {"family": "Conde", "given": "Jo\u00e3o P", "initials": "JP"}], "type": "journal article", "published": "2025-05-15", "journal": {"title": "Anal. Chim. Acta", "issn": "1873-4324", "volume": "1351", "pages": "343834", "issn-l": "0003-2670"}, "abstract": "Pathogen detection in primary care is crucial not only to identify viruses like SARS-CoV-2 but also for antibiotic-resistant bacteria. While microfluidic devices enable point-of-care diagnostics, they often lack sufficient sensitivity. On-chip isothermal amplification techniques, such as padlock probing-based rolling circle amplification (PLP-RCA), can enhance specificity and sensitivity while keeping device complexity low. However, integrating PLP-RCA on-chip requires precise optimization of enzyme concentrations, flow conditions, and target capture to achieve its full potential.\n\nThis study demonstrates a microfluidic RCA assay using porous agarose microbeads as a solid-phase capture, packed inside a microfluidic device. Various target capture strategies were systematically compared and quantitatively investigated, progressing from single-stranded synthetic DNA oligonucleotides to double-stranded Staphylococcus aureus genomic DNA. The best strategy for double-stranded Staphylococcus aureus genomic DNA used a primer bound to the beads that capture the PLP and the target genomic DNA. The system integrates an amorphous-hydrogenated silicon (a-Si:H) thin film p-i-n photodiode and a high-pass interference filter, enabling on-chip fluorescence signal acquisition of amplicons. This integration allows for a fully functional PLP-RCA assay on-chip, along with the added merits of device portability and compatibility with clinical demands.\n\nThis study systematically evaluates single- and double-stranded target capture for on-chip PLP-RCA assays. It demonstrates the successful integration of microfluidics with a solid-phase capture medium and fluorescence detection system. The findings highlight the potential of this platform for developing sensitive, portable pathogen detection devices suited for clinical applications.\"", "doi": "10.1016/j.aca.2025.343834", "pmid": "40187866", "labels": [], "xrefs": [{"db": "pii", "key": "S0003-2670(25)00228-4"}], "notes": [], "created": "2026-08-20T06:41:27.285Z", "modified": "2026-08-20T06:41:27.324Z"}