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XXII Congresso Nazionale della Società Chimica Italiana FIRENZE, 10-15 SETTEMBRE 2006. Development of a DNA-based piezoelectric biosensor for the simultaneous detection and genotyping of high risk Human Papilloma Virus strains. D. Dell’Atti , M. Minunni, S. Tombelli, M. Mascini
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XXII Congresso Nazionale della Società Chimica Italiana FIRENZE, 10-15 SETTEMBRE 2006 Development of a DNA-based piezoelectric biosensor for the simultaneous detection and genotyping of high risk Human Papilloma Virus strains D. Dell’Atti, M. Minunni, S. Tombelli, M. Mascini Università degli Studi di Firenze, Dipartimento di Chimica A. Cavazzana, M. Zavaglia, G. Bevilacqua Università degli Studi di Pisa, Dipartimento di Oncologia, dei trapianti e delle nuove tecnologie in medicina
DNA Based Biosensors Synthesisof a DNA fragment (probe) containing the sequence of interest Immobilisationof the probe onto the solid support of the sensor Extraction of the DNA from the real sample (blood, water, food) andamplificationof the sequence of interest (thiol/dextran/streptavidin/biotinylatedprobe) Denaturation of the amplified fragment to obtain a single-stranded DNA Hybridisation of the obtained ssDNA with the immobilised probe Changes in the physicochemical parameters of the layer formed on the transducer
Instrumentation • 9.5 MHz AT-Cut quartz crystal (14mm) • Gold evaporated (42.6mm2 area) on both sides • Crystal Manufactoring (USA) Methacrylate cell QCMagic (ElbaTech, Marciana, Italy)
11-mercaptoundecanol dextran gold streptavidin biotinylated probe Immobilisationprotocol
Biosensor signals Probe immobilisation A. Hybridisation buffer Target ( 10 min) C. Hybridisation buffer D. Regenerating solution HCl 1mM ( 2*30”) Hybridisation cycles A D A Analytical datum is ∆F (Fc-Fa) B C