A DNA piezoelectric sensor has been developed for the detection of genetically modified organisms (GMOs). Single stranded DNA (ssDNA) probes were immobilised on the sensor surface of a quartz crystal microbalance (QCM) device and the hybridisation between the immobilised probe and the target complementary sequence in solution was monitored. The probe sequences were internal to the sequence of the 35S promoter (P) and Nos terminator (T), which are inserted sequences in the genome of GMOs regulating the transgene expression. Two different probe immobilisation procedures were applied: (a) a thiol-dextran procedure and (b) a thiol-derivatised probe and blocking thiol procedure. The system has been optimised using synthetic oligonucleotides, which were then applied to samples of plasmidic and genomic DNA isolated from the pBI121 plasmid, certified reference materials (CRM), and real samples amplified by the polymerase chain reaction (PCR). The analytical parameters of the sensor have been investigated (sensitivity, reproducibility, lifetime etc.). The results obtained showed that both immobilisation procedures enabled sensitive and specific detection of GMOs, providing a useful tool for screening analysis in food samples. (C) 2002 Elsevier Science B.V. All rights reserved.
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Austraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, AustriaAustraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, Austria
Vollenhofer, S
Burg, K
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Austraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, AustriaAustraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, Austria
Burg, K
Schmidt, J
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Austraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, AustriaAustraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, Austria
Schmidt, J
Kroath, H
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Austraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, AustriaAustraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, Austria
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Austraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, AustriaAustraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, Austria
Vollenhofer, S
Burg, K
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Austraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, AustriaAustraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, Austria
Burg, K
Schmidt, J
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机构:
Austraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, AustriaAustraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, Austria
Schmidt, J
Kroath, H
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Austraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, AustriaAustraian Res Ctr Siebersdorf, Biotechnol Unit, A-2444 Seibersdorf, Austria