Smartphone-readable RPA-LFA for the high-sensitivity detection of Leishmania kDNA using nanophosphor reporters
Year: 2023
Type: article
Abstract: Early diagnosis of infectious diseases improves outcomes by enabling earlier delivery of effective treatment, and helps prevent further transmission by undiagnosed persons. We demonstrated a proof-of-concept assay combining isothermal amplification and lateral flow assay (LFA) for early diagnosis of cutaneous leishmaniasis, a vector-borne infectious disease that affects ca. 700,000 to 1.2 million people annually. Conventional molecular diagnostic techniques based on polymerase chain reaction (PCR) require complex apparatus for temperature cycling. Recombinase polymerase amplification (RPA) is an isothermal DNA amplification method that has shown promise for use in low-resource settings. Combined with lateral flow assay as the readout, RPA-LFA can be used as a point-of-care diagnostic tool with high sensitivity and specificity, but reagent costs can be problematic. In this work, we developed a highly-sensitive smartphone-based RPA-LFA for the detection of Leishmania panamensis DNA using blue-emitting [(Sr0.625Ba0.375)1.96Eu0.01Dy0.03]MgSi2O7 (SBMSO) persistent luminescent nanophosphors as LFA reporters. The greater detectability of nanophosphors allows the use of a reduced volume of RPA reagents, potentially reducing the cost of RPA-LFA. The limit of detection (LOD) of RPA with gold nanoparticle-based LFA readout is estimated at 1 parasite per reaction, but LOD can be 100-fold better, 0.01 parasites per reaction, for LFA based on SBMSO. This approach may be useful for sensitive and cost-effective point-of-care diagnosis and contribute to improved clinical and economic outcomes, especially in resource-limited settings.
Source: PLoS neglected tropical diseases
Language: English
Subfield: Biomedical Engineering
Field: Engineering
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $2,495
Funders: U.S. Department of Health and Human Services, Consortia for Improving Medicine with Innovation and Technology, National Institutes of Health, Centers for Disease Control and Prevention, National Institute of Allergy and Infectious Diseases, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institute of Biomedical Imaging and Bioengineering
Awards: G1001581127, WESTERN GULF CENTER OF EXCELLENCE FOR VECTOR-BORNE DISEASES, G1555212569, Smartphone-based POC Testing for HIV Using Glowstick Chemistry, Point of Care Technology Research Center in Primary Care - ITAP, G211600740, G2263952496, G2287028825, G2473118667, Western Gulf Center of Excellence for Vector-Borne Diseases, G5744357232, G6527333995, Novel Point of Care assays for Urinary Diagnostics of Nephritis, G7380516210, G9000134163, A smartphone-based point-of-care test for detection of Chlamydia trachomatis, G985056232
OpenAlex ID: W4382931758