Sensitive and high throughput quantification of abscisic acid based on quantitative real time immuno-PCR

文献类型： 外文期刊

第一作者： Li, Wei

作者： Li, Wei;Huang, Zhigang;Wang, Ruozhong;Luo, Weigui;Liu, Qing;Tong, Jianhua;Xiao, Langtao;Su, Yi;Huang, Zhigang;Wang, Ruozhong;Liu, Qing;Xiao, Langtao;Li, Wei

作者机构：

关键词： qIPCR; ABA; Biotin; Avidin

期刊名称：PLANT METHODS （ 影响因子：4.993； 五年影响因子：5.312 ）

ISSN： 1746-4811

年卷期： 2018 年 14 卷

页码：

收录情况： SCI

摘要： Abscisic acid (ABA) functions as a stress phytohormone in many growth and developmental processes in plants. The ultra-sensitive determination of ABA would help to better understand its vital roles and action mechanisms. We report a new sensitive and high throughput quantitative real time immuno-PCR (qIPCR) method based on biotin-avidin linkage system for ABA determination in plants. ABA monoclonal antibody (McAb) coated on the inner surface of PCR well pretreated with glutaraldehyde. The pre-prepared probe complex, including biotinylated McAb, biotinylated DNA and streptavidin linker, was convenient for high throughput operations. Finally, probe DNA was quantified by real-time PCR. The detectable ranges were from 10 to 40 ng/L with a limit of detection (LOD) of 2.5 fg. ABA contents in plant sample were simultaneously analyzed using LC-MS/MS to validate the qIPCR method. The results showed that qIPCR method has good specificity and repeatability with a recovery rate of 96.9%. The qIPCR method is highly sensitive for ABA quantification for actual plant samples with an advantage of using crude extracts instead of intensively purified samples.

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