Innovation
Focusing on the research and development and application of isothermal nucleic acid amplification technology
Study on the Development of a Detection Method for Non-Tuberculous Mycobacteria Based on Recombinase-Aided Isothermal Amplification—Zhejiang Inspection and Quarantine & Hangzhou Center for Disease Control and Prevention
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Abstract: Objective To establish a simple and rapid nucleic acid detection method for non-tuberculous mycobacteria under constant-temperature conditions, providing reference data for clinical diagnostics. Methods Specific primers targeting three non-tuberculous mycobacteria—Mycobacterium marinum, Mycobacterium abscessus, and Mycobacterium kansasii—were designed. A recombinase-mediated isothermal amplification technique was employed to develop a highly efficient and rapid assay for the detection of non-tuberculous mycobacteria. Results The recombinase-mediated isothermal amplification fluorescence reaction performed at 39°C yielded detectable results within approximately 20 minutes. The detection limit of this assay for the three non-tuberculous mycobacteria reached as low as 10 copies/L. No cross-reactivity was observed with Mycobacterium tuberculosis or other clinically common pathogens, resulting in a specificity of 100.0%. The assay demonstrated excellent sensitivity and specificity in detecting clinically relevant non-tuberculous mycobacterial strains. Conclusion This method provides a specific, sensitive, and rapid detection approach for non-tuberculous mycobacteria that can be readily implemented in simple laboratory settings.
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