期刊
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
卷 186, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.jpba.2020.113333
关键词
Mycobacterium tuberculosis; Mini-potentiostat; Graphene; Screen-printed electrode; LAMP-EC
资金
- National Research Council of Thailand [GRA-BT-2559-2058-TH]
- Drug Resistant Tuberculosis Research Fund under the Patronage to Her Royal Highness Princess Galayani Vadhana Krom Luang Naradhiwas Rajanagarindra, Siriraj Foundation, Faculty of Medicine Siriraj Hospital, Mahidol University
- Thailand Research Fund [RTA6180004]
Tuberculosis (TB) is one of the most contagious and lethal infectious diseases that affects more than 10 million individuals worldwide. A lack of rapid TB diagnosis is partly responsible for its alarming spread and prevalence in many regions. To address this problem, we report a novel integrated point-of-care platform to detect a TB-causative bacterium, Mycobacterium tuberculosis (Mtb). This leverages loop-mediated isothermal amplification (LAMP) for Mtb-DNA amplification and the screen-printed graphene electrode (SPGE) for label-free electrochemical analysis of DNA amplicons. When implemented on a portable potentiostat device developed in-house, the system (LAMP-EC) offers a rapid end-point qualitative analysis of specific DNA amplicons that will be displayed as a discrete positive/negative readout on the LCD screen. Under optimized conditions, LAMP-EC showed a comparable detection limit to the previously developed LAMP assay with a lateral flow readout at 1 pg total DNA or 40 Mtb genome equivalents. This highly specific technique detected the presence of TB in all 104 blinded sputum samples with a 100% accuracy. Our technique can also easily be clinically adopted due to its affordability (similar to USD2.5/test), rapidity (<65 min turnaround time) and feasibility (lack of advanced instrumental requirement). This serves as a practical incentive, appealing to users in both high- and low-resource settings across the TB endemic regions and economic backgrounds. (C) 2020 Elsevier B.V. All rights reserved.
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