Journal
ANALYTICAL CHEMISTRY
Volume 90, Issue 11, Pages 6922-6928Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.8b01160
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Funding
- Chemical Measurement and Imaging Program at the National Science Foundation [CHE-1709372]
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Point-of-care (POC) technologies for the detection of pathogens in clinical samples are highly valued Furthermore, they are ideally suited for resource-limited settings where expensive and sophisticated laboratory equipment may not be readily available. In this study, a rapid method based on solid-phase microextraction (SPME) of mycobacterial DNA with subsequent isothermal amplification and visual detection was developed. Direct coupling of the SPME desorption solution (1 M NaCl) to the isothermal reaction system was achieved to circumvent dilution steps and improve detection limits. Using this method, DNA was preconcentrated from lysed mycobacteria in just 2 min, subjected to isothermal multiple-self-matching-initiated amplification (IMSA), and the amplicons were detected visually. With a total analysis times of less than 2 h, the optimized method was capable of extracting and visually detecting mycobacterial DNA from artificial sputum samples containing clinically relevant concentrations of mycobacteria (10(7) colony forming units/mL), demonstrating its potential for future POC applications.
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