CHA结合DNAzyme用于沙门氏菌核酸的可视化检测  

Visual Detection of Salmonella DNA Fragments by CHA Combined DNAzyme

作　　者：卜胜君 王奎宇 宋广萍 英娜[2] 齐炳尧 李忠义[2] 汤鑫[2] 万家余[2] 张红梅[1] 

机构地区：[1]黑龙江八一农垦大学,黑龙江大庆163319 [2]军事医学科学院军事兽医研究所,长春130122

出　　处：《东北农业科学》2017年第3期44-49,共6页Journal of Northeast Agricultural Sciences

基　　金：吉林省科技发展计划(20150101105JC)

摘　　要：为建立简洁、灵敏、快速的食源性沙门氏菌核酸可视化检测方法,试验采用靶标催化发夹装置(Catalyzed hairpin assembly,CHA)结合功能性核酸DNAzyme的方法,设计沙门氏菌核酸发夹型抓取探针(Capture probe,CP),CP结合靶标并启动H1、H2引发CHA反应,形成DNAzyme结构并发生显色反应。试验中对CP、H1、H2最佳浓度及缓冲液pH进行优化,对灵敏度与特异性进行分析。结果表明:该方法对沙门氏菌核酸片段的可视化检测具有高特异性,灵敏度达到3.9pM,且存在一定的线性关系。说明此方法的建立可为食源性病菌可视化检测奠定相应基础。A target catalyzed hairpin assembly combined with DNAzyme was adopted to establish a simple and sen-sitive method for visual detection of salmonella specific DNA fragment. Capture probe was designed by specific sal-monella DNA segments. When target was present, capture probe opened and started the CHA reaction. H1-H2 dip-loid DNAzyme structure was formed and caused chromogenic reaction. The experiment was carried out to select opti-mal content of CP, H1, H2, and pH of buffer solution, and analyze the sensitivity and specificity. The results showedthat the system had a good generality and met the requirements of the naked eye visual detection. The detect limit ofthis method was 3.9 pM and there was a linear correlation. Setting up of this method laid basis for visual detection offood-borne bacteria.

关 键 词：沙门氏菌 特异性 16s RRNA CHA DNAZYME 可视化检测 

分 类 号：Q936[生物学—微生物学]