Universal LNA Probe-Mediated Multiplex Droplet Digital Polymerase Chain Reaction for Ultrasensitive and Accurate Quantitative Analysis of Genetically Modified Organisms

文献类型： 外文期刊

第一作者： Yang, Litao

作者： Yang, Litao;Chen, Yi;Li, Rong;Xu, Wenting;Zhang, Dabing;Cui, Jinjie;Zhang, Xiujie

作者机构： Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Joint Int Res Lab Metab & Dev Sci, Shanghai 200240, Peoples R China;Chinese Acad Agr Sci, Inst Cotton Res, State Key Lab Cotton Biol, Anyang 455000, Henan, Peoples R China;Minist Agr Peoples Republ China, Dev Ctr Sci & Technol, Beijing 100025, Peoples R China

关键词： universal LNA probe; ULNA-ddPCR; nucleic acid quantification; GM rice

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 2020影响因子：5.279； 五年影响因子：5.269 ）

ISSN： 0021-8561

年卷期： 2021 年 69 卷 5 期

页码：

收录情况： SCI

摘要： Multiplex and high-throughput assays are becoming the main trends in the development of new nucleic acid detection and quantification methods, such as those for genetically modified organism (GMO) analysis. Here, we report a novel universal LNA probe-mediated droplet digital polymerase chain reaction (PCR) method (ULNA-ddPCR) for multiple DNA target quantification in GMOs. In ULNA-ddPCR, only one universal LNA probe is used for multiple DNA targets instead of using one to one TaqMan probe. The specificity, sensitivity, dynamic range, and accuracy of the ULNA-ddPCR method are determined by employing GM rice analysis as an example. Simplex and triplex ULNA-ddPCR assays for three GM rice events, T2A-1, T1C-19, and G6H1, are established and evaluated. All results indicate that the developed simplex and triplex ULNA-ddPCR assays are suitable for quantitative analysis of GM rice events with high sensitivity, accuracy, and low cost. The ULNA-ddPCR method also has the potential for multiple DNA target quantification in other research fields.

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