Quartz crystal microbalance (QCM) affinity biosensor for genetically modified organisms (GMOs) detection
Quartz crystal microbalance (QCM) affinity biosensor for genetically modified organisms (GMOs) detection
复制标题
DOI：
10.1016/s0956-5663(02)00166-5
复制
发表时间：
2003-03-01
影响因子：
12.6
通讯作者：
Mascini, M
中科院分区：
文献类型：
--
作者：
Mannelli, I;Minunni, M;Mascini, M
研制了一种用于转基因生物检测的DNA压电传感器。将单链DNA（ssDNA）探针固定在石英晶体微天平（QCM）装置的传感器表面上，并监测固定化探针与溶液中的靶互补序列之间的杂交。探针序列位于35 S启动子（P）和Nos终止子（T）序列的内部，它们是调节转基因表达的GMO基因组中的插入序列。应用两种不同的探针固定程序：（a）硫醇-葡聚糖程序和（B）硫醇衍生化探针和封闭硫醇程序。使用合成寡核苷酸对系统进行了优化，然后将其应用于从pBI 121质粒分离的质粒和基因组DNA样品、经认证的参考物质（CRM）和通过聚合酶链反应（PCR）扩增的真实的样品。传感器的分析参数进行了研究（灵敏度，再现性，寿命等）。结果显示，两种固定化方法均能灵敏而特异地检测基因改造生物，为食品样品的筛选分析提供了有用的工具。(C)2002 Elsevier Science B. V.保留所有权利。
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.