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Microchimica Acta

, 185:314 | Cite as

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

  • Maria Magiati
  • Areti Sevastou
  • Despina P. Kalogianni
Original Paper

Abstract

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.

Graphical abstract

Schematic presentation of a fluorometric lateral flow assay based on fluorescein and phycoerythrin fluorescent labels for the detection of single-stranded (ssDNA) and double-stranded DNA (dsDNA) sequences and using a digital camera readout. SA: streptavidin, BSA: Bovine Serum Albumin, B: biotin, FITC: fluorescein isothiocyanate, PE: phycoerythrin, TZ: test zone, CZ: control zone.

Keywords

Biosensor DNA PCR Fluorescence Phycoerythrin Fluorescein isothiocyanate Strip 

Notes

Acknowledgements

We acknowledge the assistance of Dr. Sotirios Tragoulias with image acquisition.

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_2856_MOESM1_ESM.docx (207 kb)
ESM 1 (DOCX 207 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of ChemistryUniversity of PatrasPatrasGreece

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