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

机译:石英晶体微天平(QCM)亲和力生物传感器,用于转基因生物(GMO)检测

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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. [References: 34]
机译:已开发出一种DNA压电传感器,用于检测转基因生物(GMO)。将单链DNA(ssDNA)探针固定在石英晶体微量天平(QCM)设备的传感器表面,并监测溶液中固定探针与目标互补序列之间的杂交。探针序列在35S启动子(P)和Nos终止子(T)的序列内部,它们是调节转基因表达的GMOs基因组中的插入序列。应用了两种不同的探针固定程序:(a)硫醇-葡聚糖程序,(b)硫醇衍生的探针和封闭硫醇程序。该系统已使用合成寡核苷酸进行了优化,然后将其应用于从pBI121质粒分离的质粒和基因组DNA样品,经认证的参考材料(CRM)和通过聚合酶链反应(PCR)扩增的真实样品。已经研究了传感器的分析参数(灵敏度,再现性,寿命等)。获得的结果表明,两种固定方法均可实现转基因生物的灵敏和特异检测,为筛选食品样品中的分析提供了有用的工具。 (C)2002 Elsevier Science B.V.保留所有权利。 [参考:34]

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