A fluorometric lateral flow assay for visual detection of nucleic acids using a digital camera readout

被引：0
作者：
Maria Magiati
Areti Sevastou
Despina P. Kalogianni
机构：
[1] University of Patras,Department of Chemistry
来源：
Microchimica Acta | 2018年 / 185卷
关键词：
Biosensor; DNA; PCR; Fluorescence; Phycoerythrin; Fluorescein isothiocyanate; Strip;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
A fluorometric lateral flow assay has been developed for the detection of nucleic acids. The fluorophores phycoerythrin (PE) and fluorescein isothiocyanate (FITC) were used as labels, while a common digital camera and a colored vinyl-sheet, acting as a cut-off optical filter, are used for fluorescence imaging. After DNA amplification by polymerase chain reaction (PCR), the biotinylated PCR product is hybridized to its complementary probe that carries a poly(dA) tail at 3΄ edge and then applied to the lateral flow strip. The hybrids are captured to the test zone of the strip by immobilized poly(dT) sequences and detected by streptavidin-fluorescein and streptavidin-phycoerythrin conjugates, through streptavidin-biotin interaction. The assay is widely applicable, simple, cost-effective, and offers a large multiplexing potential. Its performance is comparable to assays based on the use of streptavidin-gold nanoparticles conjugates. As low as 7.8 fmol of a ssDNA and 12.5 fmol of an amplified dsDNA target were detectable.
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