Quantification of genetically modified soybean by quenching probe polymerase chain reaction.
Quantification of genetically modified soybean by quenching probe polymerase chain reaction.
复制标题
通过淬灭探针聚合酶链反应对转基因大豆进行定量。
DOI：
10.1021/jf048031r
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发表时间：
2005
影响因子：
6.1
通讯作者：
T. Kanagawa
中科院分区：
文献类型：
--
作者：
Hidenori Tani;N. Noda;Kazutaka Yamada;S. Kurata;S. Tsuneda;A. Hirata;T. Kanagawa
猝灭探针（QProbe）聚合酶链反应（PCR）是一种简单、经济的实时PCR检测方法，与其他实时PCR检测方法如TaqMan检测方法相比。采用QProbe-PCR技术对转基因大豆（抗农达大豆）进行定量分析。我们设计了Le1（大豆内源基因）和RRS（重组基因）的事件特异性q探针，并对含有0.1、0.5、1、2和5%转基因大豆的认证标准物质进行了定量。同样的样品也采用TaqMan法，并对结果进行比较。QProbe-PCR的准确性与TaqMan法相近。当转基因大豆含量大于等于0.5%时，QProbe-PCR的相对标准偏差小于20%。QProbe-PCR足够灵敏，可以监测标记系统，并具有可接受的准确性和精密度。
Quenching probe (QProbe) polymerase chain reaction (PCR) is a simple and cost-effective real-time PCR assay in comparison with other real-time PCR assays such as the TaqMan assay. We used QProbe-PCR to quantify genetically modified (GM) soybean (Roundup Ready soybean). We designed event-specific QProbes for Le1 (soy endogenous gene) and RRS (recombinant gene), and we quantified certified reference materials containing 0.1, 0.5, 1, 2, and 5% GM soybean. The TaqMan assay was also applied to the same samples, and the results were compared. The accuracy of QProbe-PCR was similar to that of TaqMan assay. When GM soybean content was 0.5% or more, the relative standard deviations of QProbe-PCR were less than 20%. QProbe-PCR is sensitive enough to monitor labeling systems and has acceptable levels of accuracy and precision.