基于链置换扩增反应与DNA银簇的免标记核酸检测方法  

A Label-Free Detection Method for Nucleic Acid Based on Strand Displacement Amplification Reaction and DNA Silver Nanoclusters

作　　者：张瑶[1] 成玉梁[1] 谢云飞 姚卫蓉[1] 郭亚辉 钱和[1,2] ZHANG Yao;CHENG Yuliang;XIE Yunfei;YAO Weirong;GUO Yahui;QIAN He(School of Food Science and Technology,Jiangnan University/State Key Laboratory of Food Science and Technology,Wuxi 214122,Jiangsu,China;International Joint Laboratory of Food safety,Jiangnan University,Wuxi 214122,Jiangsu,China)

机构地区：[1]江南大学食品学院/食品科学与技术国家重点实验室,江苏无锡214122 [2]江南大学食品安全国际联合实验室,江苏无锡214122

出　　处：《武汉大学学报（理学版）》2018年第4期349-353,共5页Journal of Wuhan University:Natural Science Edition

基　　金：国家自然科学基金(31601552);江苏省重点研发计划专项资金(BE2016664)资助项目

摘　　要：提出一种基于链置换扩增反应（SDA）和DNA银簇合成的免标记核酸检测方法.该方法分为两步：第一步,模板链（TemDNA）特异性识别目标链（TarDNA）并利用链置换扩增反应扩增出银链（AgDNA）;第二步,将扩增后的AgDNA链与硝酸银合成DNA银簇,再根据产物的荧光强度实现定量检测.该方法在0~1 200nmol·L^-1浓度范围内对目标DNA具有良好的线性响应,检出限达540pmol·L^-1,且可以根据不同的目标DNA修改模板序列.A novel label-free method to detect target DNA based on strand displacement amplification（SDA）and synthesis of silver nanoclusters（AgNCs）was developed.This method consisted of two steps：1）the target DNA（TarDNA）was recognized specially by the template DNA（TemDNA）and then amplified a lot of Ag-strand（AgDNA）using strand displacement amplification reaction（SDA）;2）amplified AgDNA was used to synthesize DNA silver nanoclusters with silver nitrate.Then the synthesized product could be quantitatively detected according to the fluorescence intensity.This detection shows a good liner response to the target DNA in the range of 0-1 200 nmol·L^-1,and the detection limit reaches 540 pmol·L^-1.Furthermore,the template DNA sequence can be modified on the basis of different target DNA sequence.

关 键 词：链置换扩增反应 银纳米簇 荧光强度 DNA聚合酶 限制性内切酶 

分 类 号：Q503[生物学—生物化学] O069[理学]