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化脓性链球菌靶向DNA适配体在熟鸡肉中的筛选、表征及应用

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

作者信息

Huang Yukun, Wang Xin, Duan Nuo, Xia Yu, Wang Zhouping, Che Zhenming, Wang Lijun, Yang Xiao, Chen Xianggui

机构信息

Key Laboratory of Grain and Oil Processing and Food Safety of Sichuan Province, School of Food and Bioengineering, Xihua University, Chengdu, 610039, China.

State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China; Shandong Gold Mining (Laizhou) Co., Ltd., Sanshan Island Gold Mine, Laizhou, 261442, China.

出版信息

Anal Biochem. 2018 Jun 15;551:37-42. doi: 10.1016/j.ab.2018.04.015. Epub 2018 Apr 23.

Abstract

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) value of 44 ± 5 nmol L 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. 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.

摘要

筛选并鉴定了一种抗化脓性链球菌的适配体,并基于所报道的适配体建立了一种荧光方法,用于检测熟鸡肉中的化脓性链球菌。通过体外十二轮全菌SELEX(指数富集配体系统进化)筛选,获得了一组与化脓性链球菌全细胞结合的适配体,其中适配体S-12的解离常数(K)值低至44±5 nmol/L。通过基于荧光共振能量转移的分析方法,利用氧化石墨烯对熟鸡肉样品中的化脓性链球菌进行基于适配体的定量分析,检测限为70 cfu/mL。所筛选的适配体对化脓性链球菌具有亲和力和选择性;此外,基于所筛选的适配体作为分子识别元件,可开发更多用于化脓性链球菌创新检测的生物传感器。

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