Leimanis Serge, Hernández Marta, Fernández Sophie, Boyer Francine, Burns Malcolm, Bruderer Shirin, Glouden Thomas, Harris Neil, Kaeppeli Othmar, Philipp Patrick, Pla Maria, Puigdomènech Pere, Vaitilingom Marc, Bertheau Yves, Remacle José
Unité de Recherche en Biologie Cellulaire (URBC), Faculté Universitaire Notre Dame de la Paix, Namur, Belgium.
Plant Mol Biol. 2006 May;61(1-2):123-39. doi: 10.1007/s11103-005-6173-4.
A multiplex DNA microarray chip was developed for simultaneous identification of nine genetically modified organisms (GMOs), five plant species and three GMO screening elements, i.e. the 35S promoter, the nos terminator and the nptII gene. The chips also include several controls, such as that for the possible presence of CaMV. The on-chip detection was performed directly with PCR amplified products. Particular emphasis was placed on the reduction of the number of PCR reactions required and on the number of primers present per amplification tube. The targets were biotin labelled and the arrays were detected using a colorimetric methodology. Specificity was provided by specific capture probes designed for each GMO and for the common screening elements. The sensitivity of the assay was tested by experiments carried out in five different laboratories. The limit of detection was lower than 0.3% GMO for all tests and in general around 0.1% for most GMOs. The chip detection system complies with the requirements of current EU regulations and other countries where thresholds are established for the labelling of GMO.
开发了一种多重DNA微阵列芯片,用于同时鉴定9种转基因生物(GMO)、5种植物物种和3种GMO筛选元件,即35S启动子、nos终止子和nptII基因。这些芯片还包括几个对照,例如用于检测是否可能存在花椰菜花叶病毒(CaMV)的对照。芯片上的检测直接使用PCR扩增产物进行。特别强调减少所需的PCR反应数量以及每个扩增管中引物的数量。靶标用生物素标记,阵列使用比色法进行检测。通过为每种GMO和常见筛选元件设计的特异性捕获探针提供特异性。该检测方法的灵敏度在五个不同实验室进行的实验中进行了测试。所有测试的检测限均低于0.3% GMO,大多数GMO的检测限通常约为0.1%。芯片检测系统符合欧盟现行法规以及其他为GMO标签设定阈值的国家的要求。