本文已被：浏览 8943次 下载 3876次
投稿时间：2016-08-25
投稿时间：2016-08-25
中文摘要: 建立改良环介导等温扩增（loop-mediated isothermal amplification，LAMP）检测酸土脂环酸芽胞杆菌的方法。以 酸土脂环酸芽胞杆菌的 16S rRNA 基因保守区域设计 4 条特异性引物，优化反应体系，通过荧光曲线、琼脂糖凝胶电泳 和白色沉淀判定扩增结果。同时，对 LAMP 引物特异性、灵敏度、检出限进行研究。LAMP 法在 61 ℃，60 min 内完成酸土 脂环酸芽胞杆菌的检测。2 株酸土脂环酸芽胞杆菌呈阳性结果， 17 株非酸土脂环酸芽胞杆菌呈阴性结果， 表明该种检 测方法具有高特异性。检测纯菌灵敏度为 7.2 CFU/mL，对人工污染苹果汁样品中酸土脂环酸芽胞杆菌的检出限是18 CFU/mL。LAMP 法检测酸土脂环酸芽胞杆有操作简单、灵敏度高、特异性强等优点，应用前景广阔。
Abstract:Development of loop-mediated isothermal amplification method was established to detect for Alicy－ clobacillus acidoterrestris（A. acidoterrestris）．Taking A. acidoterrestris 16S rRNA gene specific sequence as the target sequence，the primer sequence was designed. The reaction conditions were optimized and results observed by fluorescence curve， agarose gel electrophoresis and turbidity. The specificity， sensitivity and detection limit were detected by this method.The detection could be complished at 61 °C within 60 min by LAMP. The speci－ ficity of the assay was confirmed using 2 A. acidoterrestris strains and 17 non-A. acidoterrestris strains. The de－ tection sensitivity and detection limit were 7.2 CFU/mL， 18 CFU/mL， respectively.The advantages of simple op－ eration， high specificity and sensitivity of LAMP maked it to apply in future.
keywords: loop -mediated isothermal amplification （LAMP） Alicyclobacillus acidoterrestris 16S rRNA detection
文章编号：201710038 中图分类号： 文献标志码：A
基金项目:
| Author Name | Affiliation |
| YANG Yue， ZHANG Yun-zhe， ZHANG Xian-zhou， MA Xiao-yan， ZHANG Wei* | College of Food Science and Technology， Agricultural University of Hebei，Baoding 071000， Hebei， China |
引用文本：
津ICP备20000603号-1