Gold Nanoparticle-Enhanced Recombinase Polymerase Amplification for Rapid Visual Detection of Mycobacterium tuberculosis
1
Issued Date
2025-09-01
Resource Type
eISSN
20796374
Scopus ID
2-s2.0-105017186028
Pubmed ID
41002347
Journal Title
Biosensors
Volume
15
Issue
9
Rights Holder(s)
SCOPUS
Bibliographic Citation
Biosensors Vol.15 No.9 (2025)
Suggested Citation
Saikaew S., Sangboonruang S., Pongsararuk R., Srilohasin P., Butr-Indr B., Intorasoot S., Phunpae P., Tharinjaroen C.S., Arunothong S., Panyasit W., Chaiprasert A., Tragoolpua K., Wattananandkul U. Gold Nanoparticle-Enhanced Recombinase Polymerase Amplification for Rapid Visual Detection of Mycobacterium tuberculosis. Biosensors Vol.15 No.9 (2025). doi:10.3390/bios15090607 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/112438
Title
Gold Nanoparticle-Enhanced Recombinase Polymerase Amplification for Rapid Visual Detection of Mycobacterium tuberculosis
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Corresponding Author(s)
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Abstract
Tuberculosis (TB) remains a major global health challenge, particularly in resource-limited settings where access to rapid and reliable diagnostics is limited. Conventional diagnostic methods, such as smear microscopy and culture, are either time-consuming or lack adequate sensitivity. This study optimized recombinase polymerase amplification (RPA) using 16 primer combinations targeting IS6110 highly specific to the Mycobacterium tuberculosis complex (MTC). A novel naked-eye assay, TB-GoldDx, was developed by integrating RPA combined with gold nanoparticles (AuNPs), enabling equipment-free diagnostics. TB-GoldDx demonstrated a detection limit of 0.001 ng of MTB H37Rv DNA (~210 bacilli) per 25 µL reaction. Among 100 bacterial strains, it achieved 95.83% sensitivity and 100% specificity among 100 bacterial strains, comprising 72 MTB isolates and 28 nontuberculous bacterial species. In 140 sputum samples, the assay showed 81.43% sensitivity and 58.57% specificity versus acid-fast bacilli (AFB) smear microscopy, with sensitivity improving to 95.45% in high-load AFB 3+ specimens. Compared to a commercial line probe assay (LPA), TB-GoldDx exhibited slightly higher sensitivity (84.78% vs. 82.61%) but lower specificity (54.05% vs. 78.38%). Delivering rapid, visual results in under an hour, TB-GoldDx offers a low-cost, easily deployable solution for point-of-care tuberculosis detection, especially in underserved regions, reinforcing global End TB efforts.