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 strategy1,2 to detect point mutations in non amplified genomic DNA3. 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 β-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 β-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. 1. D’Agata R.; Corradini R.; Grasso G.; Marchelli R.; Spoto G. ChemBioChem 2008, 9, 2067-2070. 2. D’Agata R.; Corradini R.; Ferretti C.; Zanoli L.; Gatti M.; Marchelli R.; Spoto G. Biosensensors & Bioelectronics 2010, 25, 2095-2100. 3. D’Agata R., Breveglieri G., Zanoli L.M., Borgatti M., Spoto G., Gambari R. Analytical Chemistry 2011, DOI: 10.1021/ac2021932.

Direct detection of point mutations in non-amplified human genomic DNA by nanoparticle-enhanced surface plasmon resonance imaging

D’Agata R;SPOTO, Giuseppe;
2011

Abstract

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 strategy1,2 to detect point mutations in non amplified genomic DNA3. 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 β-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 β-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. 1. D’Agata R.; Corradini R.; Grasso G.; Marchelli R.; Spoto G. ChemBioChem 2008, 9, 2067-2070. 2. D’Agata R.; Corradini R.; Ferretti C.; Zanoli L.; Gatti M.; Marchelli R.; Spoto G. Biosensensors & Bioelectronics 2010, 25, 2095-2100. 3. D’Agata R., Breveglieri G., Zanoli L.M., Borgatti M., Spoto G., Gambari R. Analytical Chemistry 2011, DOI: 10.1021/ac2021932.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/20.500.11769/109176
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