One-tube smart genetic testing via coupling isothermal amplification and three-way nucleic acid circuit to glucometers
| Autor: | Baiyang Lu, Bingling Li, Lulu Guo, Yidan Tang, Qing Dong, Yan Du |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Loop-mediated isothermal amplification
Multi-analysis 02 engineering and technology Transduction (psychology) 01 natural sciences Biochemistry Signal Article Maltose-Binding Proteins Analytical Chemistry Isothermal amplification Glucometer Cronobacter sakazakii Nucleic Acids Escherichia coli Environmental Chemistry Multiplex Genetic Testing Spectroscopy Nucleic acid circuitry biology Portable detection Chemistry 010401 analytical chemistry 021001 nanoscience & nanotechnology biology.organism_classification 0104 chemical sciences Coupling (computer programming) Nucleic acid 0210 nano-technology Biological system Nucleic Acid Amplification Techniques Sensitivity (electronics) Blood Chemical Analysis Bacterial Outer Membrane Proteins |
| Zdroj: | Analytica Chimica Acta |
| ISSN: | 0003-2670 |
| DOI: | 10.1016/j.aca.2020.01.068 |
| Popis: | Urgent demand for portable diagnosis has promoted a new sensing strategy that uses personal glucometer (PGM) to detect non-glucose targets. Even though great progresses have been achieved in terms of target range and sensing principle, issues such as low final signal-to-background ratio and hard-to-realize one-tube smart analysis still exist and challenge real-world applications in gene detection. Here we propose a practical solution via coupling isothermal amplification (i.e. LAMP) and three-way amplifiable catalytic hairpin assembly (i.e. CHA) to a PGM. It allows direct transduction from genomic information to commercial portable devices with all of ultra-high sensitivity, specificity and enhanced signal-to-noise ratio. Compared with previous report without signal amplification, the introduction of CHA has successfully improved the signal amplitude by at least 12.5 folds. More importantly, through importing an effective three-way junction based transduction, we also innovatively develop a one-tube logical or multiplex analysis strategy in PGM based detection. Totally four situations of two foodborne bacteria genes, in Cronobacter sakazakii (ompA) and Escherichia coli (malB), could be directly readout using the final PGM signals, with the lowest detection amount down to less than 100 molecular copies (6.6 × 10−18 M). It is believed such a LAMP-CHA-PGM method has been already sensitive, specific, and of great potential for practically portable gene diagnostics. Graphical abstract We report a practical genetic testing method via coupling isothermal amplification and three-way nucleic acid circuit to a personal glucometer. It allows direct transduction from genomic information to commercial portable devices with all of ultra-high sensitivity, specificity and enhanced signal-to-noise ratio. Logical or multiplex analysis can also be realized in one-tube, with only one signal probe, the glucose.Image 1 Highlights • CHA circuit improves the signal-to-background ratio by at least 3.5 folds. • Misreading frequently induced by signal errors or noises is efficiently reduced. • The detection limit for pathogen gene is down to 100 molecular copies. • The method for the first time enables one-tube multiplex assay for PGM based detection. • Four situations for two pathogen genes can be are directly distinguished using the PGM. |
| Databáze: | OpenAIRE |
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