Screening aptamers for serine β-lactamase-expressing bacteria with Precision-SELEX

被引：8

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

来源：

TALANTA | 2021年 / 224卷

基金：

中国国家自然科学基金;

关键词：

Aptamers; Precision-SELEX; Bacteria; High-content imaging; Nanoparticle; DNA APTAMERS; CELLS; PROBE;

D O I：

10.1016/j.talanta.2020.121750

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

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.

引用

页数：11