Quartz crystal microbalance (QCM) affinity biosensor for genetically modified organisms (GMOs) detection

被引:170
作者
Mannelli, I [1 ]
Minunni, M [1 ]
Tombelli, S [1 ]
Mascini, M [1 ]
机构
[1] Univ Florence, Dipartimento Chim, I-50019 Sesto Fiorentino, Italy
关键词
QCM; GMO; immobilisation procedure;
D O I
10.1016/S0956-5663(02)00166-5
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
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.
引用
收藏
页码:129 / 140
页数:12
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