Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University , Shanghai 200240, China.
Anal Chem. 2014 Jan 21;86(2):1269-76. doi: 10.1021/ac403630a. Epub 2014 Jan 9.
The monitoring of genetically modified organisms (GMOs) is a primary step of GMO regulation. However, there is presently a lack of effective and high-throughput methodologies for specifically and sensitively monitoring most of the commercialized GMOs. Herein, we developed a multiplex amplification on a chip with readout on an oligo microarray (MACRO) system specifically for convenient GMO monitoring. This system is composed of a microchip for multiplex amplification and an oligo microarray for the readout of multiple amplicons, containing a total of 91 targets (18 universal elements, 20 exogenous genes, 45 events, and 8 endogenous reference genes) that covers 97.1% of all GM events that have been commercialized up to 2012. We demonstrate that the specificity of MACRO is ~100%, with a limit of detection (LOD) that is suitable for real-world applications. Moreover, the results obtained of simulated complex samples and blind samples with MACRO were 100% consistent with expectations and the results of independently performed real-time PCRs, respectively. Thus, we believe MACRO is the first system that can be applied for effectively monitoring the majority of the commercialized GMOs in a single test.
转基因生物（GMO）的监测是 GMO 监管的首要步骤。然而，目前缺乏有效和高通量的方法来专门且敏感地监测大多数商业化的 GMO。在此，我们开发了一种在芯片上进行多重扩增并在寡核苷酸微阵列上进行读取的方法（MACRO），专门用于方便的 GMO 监测。该系统由用于多重扩增的微芯片和用于读取多个扩增子的寡核苷酸微阵列组成，共包含 91 个靶标（18 个通用元件、20 个外源基因、45 个事件和 8 个内参基因），涵盖了截至 2012 年商业化的所有 GM 事件的 97.1%。我们证明 MACRO 的特异性约为 100%，检测限（LOD）适合实际应用。此外，MACRO 对模拟复杂样本和盲样的检测结果与预期结果以及独立进行的实时 PCR 结果完全一致。因此，我们认为 MACRO 是第一个可用于在单次检测中有效监测大多数商业化 GMO 的系统。
