is an opportunistic pathogenic yeast that predominantly causes invasive candidiasis. Conventional methods for detecting species are costly, take 3-5 days, and require skilled technicians. Rapid pathogen identification is important in managing invasive candidiasis infection. Here, a novel molecular diagnostic assay termed loop-mediated isothermal amplification combined with nanoparticles-based lateral flow biosensor (LAMP-LFB) was developed for rapid detection A set of six primers was designed based on the species-specific internal transcribed spacer 2 (ITS2) gene. The -LAMP results were visually reported by LFB within 2 min. Various fungal strains representing species, as well as several Gram-negative and Gram-positive bacterial species, were used to determine the analytical sensitivity and specificity of the assay. The optimal LAMP conditions were 64 °C for 40 min, with a sensitivity of 1 fg of genomic DNA template from pure cultures. No cross-reactions were obtained with non- strains. Thus, the analytical specificity of the LAMP-LFB assay was 100%. The entire procedure could be completed within 85 min, including specimen processing (40 min), isothermal reaction (40 min), and result reporting (within 2 min). In 330 clinical samples (including 30 whole blood, 100 middle segment urine, and 200 sputum samples), all positive (62/330) samples were identified by LAMP-LFB assay, and the diagnostic accuracy was 100% when compared to the traditional clinical cultural-based methods. Thus, this assay can be used as a diagnostic tool for the rapid, accurate, sensitive, low-cost and specific detection of strains, especially in resource-limited settings.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9676452 | PMC |
http://dx.doi.org/10.3389/fbioe.2022.1025083 | DOI Listing |
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