Solid-Phase Microextraction of DNA from Mycobacteria in Artificial Sputum Samples To Enable Visual Detection Using Isothermal Amplification

Analytical Chemistry
Marcelino VaronaJared L Anderson

Abstract

Point-of-care (POC) technologies for the detection of pathogens in clinical samples are highly valued due to their speed, ease of use, and cost-effectiveness. 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 (107 colony forming units/mL), demonstrating its potential for future POC applications.

References

Mar 1, 1990·Journal of Clinical Microbiology·R BoomJ van der Noordaa
Feb 1, 1986·Journal of General Microbiology·R PatelP Mounts
Jun 28, 2000·Nucleic Acids Research·T NotomiT Hase
Apr 4, 2003·International Journal of Food Microbiology·Burkhard MalornyJeffrey Hoorfar
Mar 25, 2004·Journal of Mass Spectrometry : JMS·György Vas, Károly Vékey
Jun 8, 2006·PLoS Biology·Olaf PiepenburgNiall A Armes
Aug 5, 2006·Angewandte Chemie·Jeffrey B-H TokGeorge M Dougherty
Aug 30, 2008·Analytical Chemistry·Jian WenJames P Landers
Jun 17, 2010·Journal of the American Chemical Society·Juliane SchochAndres Jäschke
Mar 8, 2011·Trends in Biotechnology·Angelika NiemzDavid S Boyle
Nov 15, 2013·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Angela M CaliendoUNKNOWN Infectious Diseases Society of America (IDSA)
Dec 18, 2013·The Lancet Infectious Diseases·Paul K DrainIngrid V Bassett
Mar 1, 2013·Global Health, Science and Practice·Amy S PiatekYaDiul Mukadi
Mar 21, 2017·Journal of Pharmaceutical and Biomedical Analysis·Marcos TasconJanusz Pawliszyn
Sep 6, 2017·Analytical Chemistry·Omprakash NachamJared L Anderson
Feb 11, 2018·Pulmonology·A L García-BasteiroG Theron

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Citations

Oct 28, 2019·Analytical and Bioanalytical Chemistry·Ashley N BowersJared L Anderson
Nov 26, 2019·Chemical Communications : Chem Comm·Haiyan DongJunyang Zhuang
May 20, 2020·Science Advances·Huan XuMing Chen
Oct 20, 2018·Analytical Chemistry·María J Trujillo-RodríguezJared L Anderson
Nov 27, 2019·Analytical Chemistry·Ling XiaGongke Li
Nov 16, 2018·Analytical Chemistry·Xiong DingChangchun Liu
Nov 21, 2021·Journal of Separation Science·Sheng PengGangfeng Ouyang

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