期刊
BIOSENSORS & BIOELECTRONICS
卷 109, 期 -, 页码 177-183出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2018.03.008
关键词
Single-nucleotide polymorphism; Loop-mediated isothermal amplification; Point-of-care optical testing; Smartphone; Phannacogenomics
类别
资金
- Generalitat Valenciana [GVA-PROMETEOII/2014/040, GRISOLIA/2014/024]
- Spanish Ministry of Economy and Competitiveness [MINECO CTQ2013-45875-R]
The genotyping of a single-nucleotide polymorphism (SNP) is addressed through methods based on loop-mediated isothermal amplification (LAMP) combined with user-friendly optical read-outs to cover the current demand for point-of-care DNA biomarker detection. The modification of primer design and reaction composition improved the assay selectivity yielding allele-specific results and reducing false-positive frequency. Furthermore, the reduced cost, ease of use and effectiveness of calorimetric detection (solution and hybridisation chip formats) were availed for the image capture by a smartphone, reching high sensitivity. In order to evaluate their discriminating capacities, LAMP-based methods were applied to human samples to genotype a SNP biomarker (rs1954787) located in the GRIK4 gene and related to the treatment response to anti-depressants drugs. Sensitive (limit of detection: 100 genomic DNA copies), reproducible ( < 15% error), fast (around 70 min) and low-cost assays were accomplished. Patient subgroups were correctly discriminated, agreeing with reference sequencing techniques. The achieved analytical performances using the developed amplification-detection principles confirmed the approach potential for point-of-care optical DNA testing.
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