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用于定量检测转基因玉米事件TC - 1507的实时聚合酶链反应检测方法的实验室间转移

Interlaboratory transfer of a real-time polymerase chain reaction assay for quantitative detection of genetically modified maize event TC-1507.

作者信息

La Paz José-Luís, García-Muniz Nora, Nadal Anna, Esteve Teresa, Puigdomènech Pere, Pla Maria

机构信息

Institut de Biologia Molecular de Barcelona, Departament de Genética Molecular, Consorci CSIC-IRTA, Jordi Girona 18-26,Barcelona, Spain.

出版信息

J AOAC Int. 2006 Sep-Oct;89(5):1347-52.

Abstract

A real-time polymerase chain reaction (QPCR) assay was developed for quantitative detection of a genetically modified (GM) maize event TC-1507 and modification to conventional PCR for qualitative purposes. Sequences 5'-flanking TC-1507 full-length insert were characterized and showed multiple rearrangements involving insert and maize chloroplast fragments. The event specificity of the TC-1507 assays was based on the detection of transgene and plant rearranged sequences found to 5' flank the insertion site. They were fully specific and exhibited a limit of detection below 10 target copies, allowing consistent detection of 0.1% GM levels. The QPCR was highly linear and efficient and proved adequate for quantification of GM contents, aiming at the fulfillment of legal requirements established in the European Union (i.e., compulsory labeling of TC-1507 levels >0.9%). It satisfactorily determined TC-1507 contents on different matrixes and was successfully transferred a different laboratory.

摘要

开发了一种实时聚合酶链反应(QPCR)检测方法用于定量检测转基因玉米事件TC - 1507,并对常规PCR进行了改进以用于定性目的。对TC - 1507全长插入片段的5'侧翼序列进行了表征,结果显示存在涉及插入片段和玉米叶绿体片段的多个重排。TC - 1507检测方法的事件特异性基于对转基因和在插入位点5'侧翼发现的植物重排序列的检测。它们具有完全特异性,检测限低于10个目标拷贝,能够一致地检测出0.1%的转基因水平。该QPCR具有高度线性和高效性,被证明足以对转基因含量进行定量,旨在满足欧盟制定的法律要求(即TC - 1507水平>0.9%时必须进行标签标注)。它能令人满意地测定不同基质上的TC - 1507含量,并成功转移到了不同实验室。

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