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Aptamer-Pendant DNA Tetrahedron Nanostructure Probe for Ultrasensitive Detection of Tetracycline by Coupling Target-Triggered Rolling Circle Amplification
ACS Applied Materials & Interfaces ( IF 8.3 ) Pub Date : 2021-04-21 , DOI: 10.1021/acsami.1c02612
Chengyi Hong 1 , Xiaoxia Zhang 1 , Sishi Ye 1 , Hongfen Yang 2 , Zhiyong Huang 1 , Dan Yang 3 , Ren Cai 4 , Weihong Tan 4
Affiliation  

Tetracycline (TET) is a broad-spectrum antibiotic, which is frequently used in the prevention and treatment of animal diseases, feed additives, and so on. However, its residue and accumulation in animal-derived foods could cause several side effects to the human body. Herein, we fabricated TET aptamer-pendant DNA tetrahedral nanostructure-functionalized magnetic beads (Apt-tet MBs) as a probe to detect TET. In the presence of target TET, DNA primer was released from Apt-tet MBs since the TET aptamer could specifically bind TET. Next, the separated DNA primer could effectively initiate rolling circle amplification (RCA) reaction and generate a long tandem single-stranded sequence. Finally, with SYBR Green I as the fluorescence dye, the fluorescence signal could be detected by detection probes through hybridizing the RCA product. Under optimal conditions, the fluorescent signal increased with the increasing target TET concentration within the 5 orders of magnitude dynamic range from 0.001 to 10 ng mL–1. The detection limit was calculated to be 0.724 pg mL–1 and the method showed high selectivity toward TET among different antibiotics. More impressively, this method was employed for TET determination in fish and honey samples. The as-obtained results were consistent with those of ELISA kits, holding great potential in the field of food analysis.

中文翻译:

适体悬垂 DNA 四面体纳米结构探针通过偶联靶触发滚环扩增超灵敏检测四环素

四环素(TET)是一种广谱抗生素,常用于动物疾病的防治、饲料添加剂等。然而,其在动物源性食品中的残留和积累会对人体产生多种副作用。在此,我们制造了 TET 适配体-悬垂 DNA 四面体纳米结构功能化磁珠(Apt-tet MBs)作为检测 TET 的探针。在目标 TET 存在的情况下,DNA 引物从 Apt-tet MB 中释放出来,因为 TET 适体可以特异性结合 TET。接下来,分离的 DNA 引物可以有效启动滚环扩增 (RCA) 反应并生成长串联单链序列。最后,以SYBR Green I为荧光染料,通过与RCA产物杂交,可通过检测探针检测荧光信号。–1 . 计算出的检测限为 0.724 pg mL –1,并且该方法对不同抗生素中的 TET 显示出高选择性。更令人印象深刻的是,该方法用于测定鱼和蜂蜜样品中的 TET。所得结果与ELISA试剂盒一致,在食品分析领域具有巨大潜力。
更新日期:2021-05-05
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