Aptamer selection and application in multivalent binding-based electrical impedance detection of inactivated H1N1 virus

被引:102
作者
Bai, Chenjun [1 ]
Lu, Zhangwei [2 ]
Jiang, Hua [3 ]
Yang, Zihua [2 ]
Liu, Xuemei [1 ]
Ding, Hongmei [1 ]
Li, Hui [1 ]
Dong, Jie [1 ]
Huang, Aixue [1 ]
Fang, Tao [1 ]
Jiang, Yongqiang [3 ]
Zhu, Lingling [1 ]
Lou, Xinhui [2 ]
Li, Shaohua [1 ]
Shao, Ningsheng [1 ]
机构
[1] Beijing Inst Basic Med Sci, Dept Biochem & Mol Biol, Beijing, Peoples R China
[2] Capital Normal Univ, Dept Chem, Beijing, Peoples R China
[3] Beijing Inst Microbiol & Epidemiol, State Key Lab Pathogen & Biosecur, Beijing, Peoples R China
关键词
Aptamer; Type A influenza H1N1 virus; Inactivated virus; Electrochemical impedance aptasensor; IN-VITRO SELECTION; DNA APTAMERS; SYSTEMATIC EVOLUTION; SANDWICH ASSAY; RNA APTAMERS; H5N1; VIRUS; MOLECULES; APTASENSOR; LIGANDS; SENSOR;
D O I
10.1016/j.bios.2018.03.047
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
The type A influenza viruses are the most virulent and variable human pathogens with epidemic or even pandemic threat. The development of sensitive, specific and safe field testing methods is in particular need and quite challenging. We report here the selection and practical utilization of the inactivated influenza virus-specific aptamers. The DNA aptamers against inactivated intact H1N1 virus particles were identified through the systematic evolution of ligands by exponential enrichment (SELEX) procedure. The discriminative aptamers and their truncated sequences showed selectively high affinity to inactive H1N1 virus and H3N2 virus with the K-d in the low nanomolar range and collective binding properties. The truncated sequences were first applied in a sandwich enzyme-linked oligonucleotide assay (ELONA) with a H1N1 detection limit (LOD, S/N = 3) of 0.3 ng/and then in an electrochemical impedance (EIS) aptasensor with more than 300 times improved LOD (0.9 pg/mu L) and the excellent selectivity over other viruses (> 100 times). Therefore the developed aptasensors represent the safer, simpler, and possibly better virus-variation adaptable means of virus diagnostics.
引用
收藏
页码:162 / 167
页数:6
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