文献类型：期刊文献

英文题名：Screening aptamers for serine β-lactamase-expressing bacteria with Precision-SELEX

作者：Yu, Fang[1];Chen, Jing[1];Wang, Zecheng[1];Yang, Huixin[2];Li, Hui[1];Jia, Wenchao[1];Xue, Shuyuan[2];Xie, Hexin[2];Xu, Danke[1]

机构：[1]Nanjing Univ, Sch Chem & Chem Engn, State Key Lab Analyt Chem Life Sci, Nanjing 210046, Jiangsu, Peoples R China;[2]East China Univ Sci & Technol, Sch Pharm, State Key Lab Bioreactor Engn, Shanghai Key Lab New Drug Design, Shanghai 200237, Peoples R China

年份：2021

卷号：224

外文期刊名：TALANTA

收录：;EI(收录号：20204609482274);WOS:【SCI-EXPANDED(收录号:WOS:000600787800009)】；

基金：We acknowledge financial support of the National Natural Science Foundation of China (21775068, 21974066, 21475060 and 21227009) and Excellent Research Program of Nanjing University (ZYJH004).

语种：英文

外文关键词：Aptamers; Precision-SELEX; Bacteria; High-content imaging; Nanoparticle

摘要：Klebsiella pneumoniae carbapenemase 2 (KPC-2) is a serine beta-lactamase that can hydrolyze almost all beta-lactam antibiotics. The drug resistant problem of bacteria expressing carbapenemases is currently a global problem, therefore, rapid and specific detection of pathogenic bacteria is urgent. In order to obtain an aptamer that can specifically recognize bacteria expressing KPC-2, we have established a method called Precision-SELEX. Precision-SELEX combined protein SELEX and bacterium SELEX. In this method, KPC-2 was used as a target protein, and Escherichia coli expressing KPC-2 (KPC-2 E. coli) was used as a target bacterium. After precision-SELEX, the same aptamer named XK-10 that can recognize KPC-2 and KPC-2 E. coli was obtained while the screening process could be shortened to 4 rounds. Dissociation equilibrium constants were calculated as 0.81 nM by SPR. In addition, XK-10 could specifically bind to KPC-2 E. coli, which was confirmed through flow cytometry and molecular Docking Simulations. The high-content imaging method could detect KPC-2 E. coli. In all, the Precision-SELEX provides an accurate and efficient method to screening aptamers for bacteria.