中文摘要：目前迫切需要快速、低成本的用于监测转基因生物的多重方法。本文介绍了一个用于多重视觉检察的、基于毛细管阵列的环介导等温扩增法（CALM），该方法是可以简单、快速地监测转基因生物。在CALM中，环介导等温扩增（LAMP）引物被预固定在毛细血管内表面。毛细管阵列的表面是疏水性的，而毛细管的表面是亲水性的，因而能够通过毛细管作用力同时将LAMP的反应混合物加载和分离到每个毛细管。在进行毛细管LAMP反应的同时，作者通过手持式的紫外线装置来对结果进行视觉检测。利用CALM，我们成功地检测到了七种常用的转基因/元件和五种具有高特异性和敏感性的植物内源参考基因。另外，现实中盲样的CALM检测结果与独立实时PCR的检测结果一致。因此，我们可以用一种单一、易操作的方法检测多元核酸，CALM将成为转基因生物监测领域的一个广泛应用的技术。
外文摘要：There is an urgent need for rapid, low-cost multiplex methodologies for the monitoring of genetically modified organisms (GMOs). Here, we report a Capillary Array-based Loop-mediated isothermal amplification for Multiplex visual detection of nucleic acids (CALM) platform for the simple and rapid monitoring of GMOs. In CALM, loop-mediated isothermal amplification (LAMP) primer sets are pre-fixed to the inner surface of capillaries. The surface of the capillary array is hydrophobic while the capillaries are hydrophilic, enabling the simultaneous loading and separation of the LAMP reaction mixtures into each capillary by capillary forces. LAMP reactions in the capillaries are then performed in parallel, and the results are visually detected by illumination with a hand-held UV device. Using CALM, we successfully detected seven frequently used transgenic genes/elements and five plant endogenous reference genes with high specificity and sensitivity. Moreover, we found that measurements of real-world blind samples by CALM are consistent with results obtained by independent real-time PCRs. Thus, with an ability to detect multiple nucleic acids in a single easy-to-operate test, we believe that CALM will become a widely applied technology in GMO monitoring.
作者：Shao, Ning; Chen, Jianwei; Hu, Jiaying; 等
作者单位：Shanghai Jiao Tong Univ
期刊名称：LAB ON A CHIP
期刊影响因子：5.586
出版年份：2017
出版刊次：3
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