Magnetic bead/gold nanoparticle double-labeled primers for electrochemical detection of isothermal amplified Leishmania DNA
De La Escosura-Muñiz, Alfredo (Institut Català de Nanociència i Nanotecnologia)
Baptista Pires, Luis Miguel (Institut Català de Nanociència i Nanotecnologia)
Serrano, Lorena (Vetgenomics)
Altet Sanahujes, Laura (Vetgenomics)
Francino, Olga (Universitat Autònoma de Barcelona. Departament de Ciència Animal i dels Aliments)
Sánchez Bonastre, Armando (Centre de Recerca en Agrigenòmica)
Merkoçi, Arben (Institut Català de Nanociència i Nanotecnologia)

Date:	2016
Abstract:	A novel methodology for the isothermal amplification of Leishmania DNA using labeled primers combined with the advantages of magnetic purification/preconcentration and the use of gold nanoparticle (AuNP) tags for the sensitive electrochemical detection of such amplified DNA is developed. Primers labeled with AuNPs and magnetic beads (MBs) are used for the first time for the isothermal amplification reaction, being the amplified product ready for the electrochemical detection. The electrocatalytic activity of the AuNP tags toward the hydrogen evolution reaction allows the rapid quantification of the DNA on screen-printed carbon electrodes. Amplified products from the blood of dogs with Leishmania (positive samples) are discriminated from those of healthy dogs (blank samples). Quantitative studies demonstrate that the optimized method allows us to detect less than one parasite per microliter of blood (8 × 10⁻³ parasites in the isothermal amplification reaction). This pioneering approach is much more sensitive than traditional methods based on real-time polymerase chain reaction (PCR), and is also more rapid, cheap, and user-friendly.
Grants:	European Commission 315653 Ministerio de Economía y Competitividad SEV-2013-0295 Ministerio de Economía y Competitividad MAT2011-25870 Agència de Gestió d'Ajuts Universitaris i de Recerca 2014/SGR-260
Rights:	Tots els drets reservats.
Language:	Anglès
Document:	Article ; recerca ; Versió sotmesa a revisió
Subject:	Electrocatalytic detection ; Gold nanoparticles ; Isothermal DNA amplification ; Leishmania ; Magnetic beads
Published in:	Small, Vol. 12, issue 2 (Jan. 2016) , p. 205-213, ISSN 1613-6829

DOI: 10.1002/smll.201502350

26 p, 1.1 MB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Experimental sciences > Catalan Institute of Nanoscience and Nanotechnology (ICN2)
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Experimental sciences > CRAG (Centre for Research in Agricultural Genomics)
Articles > Research articles
Articles > Published articles