Selection and characterization, application of a DNA aptamer targeted to Streptococcus pyogenes in cooked chicken

被引:18
作者
Huang, Yukun [1 ]
Wang, Xin [2 ,3 ]
Duan, Nuo [2 ]
Xia, Yu [2 ]
Wang, Zhouping [2 ]
Che, Zhenming [1 ]
Wang, Lijun [1 ]
Yang, Xiao [1 ]
Chen, Xianggui [1 ]
机构
[1] Xihua Univ, Sch Food & Bioengn, Key Lab Grain & Oil Proc & Food Safety Sichuan Pr, Chengdu 610039, Sichuan, Peoples R China
[2] Jiangnan Univ, Sch Food Sci & Technol, State Key Lab Food Sci & Technol, Wuxi 214122, Peoples R China
[3] Shandong Gold Min Laizhou Co Ltd, Sanshan Isl Gold Mine, Laizhou 261442, Peoples R China
关键词
Aptamer; Streptococcus pyogenes; Whole-bacterium SELEX; Cooked chicken; Graphene oxide; SELEX; IDENTIFICATION; BIOSENSOR; BINDING;
D O I
10.1016/j.ab.2018.04.015
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An aptamer against Streptococcus pyogenes was selected and identified, and a fluorescent method based on the reported aptamer was established to detect S. pyogenes in the cooked chicken. Through a twelve rounds of whole-bacterium SELEX (systematic evolution of ligands by exponential enrichment) selection in vitro, a set of aptamers binding to the whole cell of S. pyogenes were generated, harvesting a low-level dissociation constant (K-d) value of 44 +/- 5 nmol L-1 of aptamer S-12. Aptamer-based quantification of S. pyogenes in the cooked chicken sample was implemented in a fluorescence resonance energy transfer-based assay by using graphene oxide, resulting in a limit of detection of 70 cfu mL(-1). The selected aptamer showed affinity and selectivity recognizing S. pyogenes; besides, more biosensors based on the selected aptamer as a molecular recognition element could be developed in the innovative determinations of S. pyogenes.
引用
收藏
页码:37 / 42
页数:6
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