Selection of an aptamer against Muscovy duck parvovirus for highly sensitive rapid visual detection by label-free aptasensor

文献类型： 外文期刊

第一作者： Lu, Taofeng

作者： Lu, Taofeng;Ma, Qin;Yan, Wenzhuo;Wang, Yuanzhi;Zhang, Yuanyuan;Zhao, Lili;Chen, Hongyan

作者机构：

关键词： Aptamer;Label-free aptasensor;Muscovy duck parvovirus;Virus detection

期刊名称：TALANTA （ 影响因子：6.057； 五年影响因子：5.386 ）

ISSN： 0039-9140

年卷期： 2018 年 176 卷

页码：

收录情况： SCI

摘要： Muscovy duck parvovirus (MDPV) causes high mortality and morbidity in ducks. This study investigated a novel aptamer-based, label-free aptasensor detection of MDPV. In this study, we developed an ssDNA aptamer using the filtration partition and lambda exonuclease method with an affinity-based monitor and counter-screening process. After 15 rounds of SELEX (systematic evolution of ligands by exponential enrichment), the ssDNA aptamer Apt-10, which specifically bound to MDPV with high affinity (K-d = 467 nM) was successfully screened, and the aptamer was also found to be good specific to MDPV. The selected Apt-10 aptamer can be used to distinguish MDPV and goose parvovirus (GPV). Three-dimensional structural analysis of the Apt-10 aptamer indicated that it folded into a compact stem-loop motif, which was related to its high affinity. Finally, a label-free detection method based on unmodified gold nanoparticles and Apt-10 aptamer was developed for MDPV determination. The concentration of Apt-10 aptamer at 5 mu M was optimal for MDPV determination in the label free aptasensor. Excellent linearity was acquired and the lowest detection limit was 1.5 or 3 EID50 (50% egg infection dose) of MDPV, respectively, depending upon spectrophotometry or the naked eye were used. These results show the potential of the aptamer for the rapid detection of MDPV and antiviral research.

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