Sample-to-answer on molecular diagnosis of bacterial infection using integrated lab-on-a-disc
| Autor: | I.L.G. Law, Czarina C. H. Leung, H.C. Kwok, M.L. Chin, Y.K. Cheung, Patrick Kwan, Mamie Hui, Y.Y. Cheung, Ho-Pui Ho, Siu Kai Kong, Jacky Fong-Chuen Loo, Shu-Yuen Wu |
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| Rok vydání: | 2017 |
| Předmět: |
Acinetobacter baumannii
DNA Bacterial Microfluidics Biomedical Engineering Biophysics Loop-mediated isothermal amplification Biosensing Techniques 02 engineering and technology 01 natural sciences Microbiology Mycobacterium tuberculosis Sepsis Lysis buffer Electrochemistry medicine Humans Organic Chemicals Medical diagnosis biology 010401 analytical chemistry General Medicine Microfluidic Analytical Techniques 021001 nanoscience & nanotechnology biology.organism_classification DNA extraction 0104 chemical sciences Sputum medicine.symptom 0210 nano-technology Biotechnology Biomedical engineering |
| Zdroj: | Biosensors and Bioelectronics. 93:212-219 |
| ISSN: | 0956-5663 |
| DOI: | 10.1016/j.bios.2016.09.001 |
| Popis: | Sepsis by bacterial infection causes high mortality in patients in intensive care unit (ICU). Rapid identification of bacterial infection is essential to ensure early appropriate administration of antibiotics to save lives of patients, yet the present benchtop molecular diagnosis is time-consuming and labor-intensive, which limits the treatment efficiency especially when the number of samples to be tested is extensive. Therefore, we hereby report a microfluidic platform lab-on-a-disc (LOAD) to provide a sample-to-answer solution. Our LOAD customized design of microfluidic channels allows automation to mimic sequential analytical steps in benchtop environment. It relies on a simple but controllable centrifugation force for the actuation of samples and reagents. Our LOAD system performs three major functions, namely DNA extraction, isothermal DNA amplification and real-time signal detection, in a predefined sequence. The disc is self-contained for conducting sample heating with chemical lysis buffer and silica microbeads are employed for DNA extraction from clinical specimens. Molecular diagnosis of specific target bacteria DNA sequences is then performed using a real-time loop-mediated isothermal amplification (RT-LAMP) with SYTO-9 as the signal reporter. Our LOAD system capable of bacterial identification of Mycobacterium tuberculosis (TB) and Acinetobacter baumanii (Ab) with the detection limits 103cfu/mL TB in sputum and 102cfu/mL Ab in blood within 2h after sample loading. The reported LOAD based on an integrated approach should address the growing needs for rapid point-of-care medical diagnosis in ICU. |
| Databáze: | OpenAIRE |
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