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Direct Detection of Point Mutations in Nonamplified Human Genomic DNA (443 visite)

D'Agata R, Breveglieri G, Zanoli LM, Borgatti M, Spoto G, Gambari R

Analytical Chemistry (ISSN: 0003-2700), 2011 Nov 15; 83(22): 8711-8717.

Tipo di articolo: Journal Article,

Impact factor: 5.856

Impact factor a 5 anni: 5.983


Parole chiave: Biomedical Applications, Diagnostic Applications, Direct Detection, Dna Amplification, Genomic Dna, Healthy Individuals, Point Mutations, Surface Plasmon Resonance Imaging, Ultrasensitive, Ultrasensitive Detection, Polymerase Chain Reaction, Gold, Metal Nanoparticle, Beta Thalassemia, Blood, Chemistry, Genetics, Human, Sensitivity And Specificity, Surface Property, Beta-Thalassemia, Surface Properties,

Url: http://www.scopus.com/inward/record.url?eid=2-s2.0-81255144060&partnerID=40&md5=6e1a9c3626f50fee61580e3dfae10893

Ultrasensitive detection protocols not requiring polymerase chain reaction (PCR)-mediated target DNA amplification are expected to significantly improve our possibilities in several research and diagnostic applications for which minute cell quantities are available. For this reason we have tested a nanoparticle-enhanced surface plasmon resonance imaging (SPRI) sensing strategy to detect point mutations in nonamplified genomic DNA. We have used genomic DNAs, not subject to costly, time-consuming, and prone to contamination PCR-based amplification procedures, obtained from both healthy individuals and homozygous or heterozygous patients affected by beta-thalassemia, in order to demonstrate the specificity and the sensitivity of the described sensing strategy. The assay we describe is ultrasensitive and convenient. Attomolar concentrations of target genomic DNA are detected, DNAs from healthy individuals and homozygous or heterozygous patients affected by beta-thalassemia are discriminated, and only simple manipulations of the genetic samples are required before the analysis. The proposed ultrasensitive detection of DNA point mutations involved in genomic disorders possibly represents an important advantage in several biomedical applications.
*** IBB - CNR ***

Dipartimento di Scienze Chimiche, Università di Catania, Viale Andrea Doria 6, I-95125 Catania, Italy

Centro di Biotecnologie, Università di Ferrara, via Fossato di Mortara 64b, 44121 Ferrara, Italy

Scuola Superiore di Catania, C/o Dipartimento di Scienze Chimiche, Università di Catania, Viale Andrea Doria 6, Catania, Italy

Istituto Biostrutture e Bioimmagini, CNR, Viale A. Doria 6, Catania, Italy

Dipartimento di Biochimica e Biologia Molecolare, Università di Ferrara, via Fossato di Mortara 74, 44121 Ferrara, Italy
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10 Records (8 escludendo Abstract e Conferenze).
Impact factor totale: 18.739 (16.925 escludendo Abstract e Conferenze).
Impact factor a 5 anni totale: 19.355 (17.536 escludendo Abstract e Conferenze).







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