环介导恒温扩增法快速检测短小芽孢杆菌.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2011 2 1 1, 2 1 1 1 1 1 1 应琪 李太武 苏秀榕 李晔 周君 崔静 王中华 李松伟 1 2 ( , 315211; , 315110) : 短小芽孢杆菌(Bacilluspum ilus)是一种能 引起食 性疾病的腐败菌, 对其进行快速检测具有重要意义针对 短小芽孢杆菌木聚糖 ( xynA)基因, 设计了 4条特异性引物(两条内引物和两条外引物 ), 通过条件优化, 首次将一种新颖的核 酸扩增技术 环介导恒温扩增技术应用于短小芽孢杆菌的快速检测采用该技术, 63 温育 1 h 的条件下扩增短小芽孢杆 菌 DNA,琼脂糖凝胶电泳得到特异性梯度条带PCR和 LAMP的检测灵敏度分别约为 162和 16. 2拷贝每反应结果表明, 该 方法检测短小芽孢杆菌特异性强灵敏度高操作简便检测成本低, 1 h即可完成,有望发展成为快速检测短小芽孢杆菌的有 效手段 : 短小芽孢杆菌 环介导恒温扩增技术( LAM P) 木聚糖( xynA) 基因 快速检测 Establishment of Loop-mediated IsothermalAmplification M ethod for Detection ofBacillus p um ilus 1 1, 2 1 1 1 1 1 1 YingQ i LiTai u Su X iurong LiYe Zhou Jun Cui Jing W ang Zhonghua Li Song ei 1 ( Faculty of Lfie Science and Biotechnology, N ingbo University,N ingbo 315211; 2 N ingbo C ity College of Vocational echnology,N ingbo 315110) Abstrac:t Bacilluspum ilus has been considered as one of themost mi portant spoilage bacterial hich also causes foodborne il-l ness. A set of four prmi ers, t o outer and t o inner prmi ers, as designed specifically to recognize the xylanase ( xynA ) gene ofB. pum ilus. After the optmi ization of loop-mediated isothermal amplification ( LAMP) reaction m ix, the LAMP method as applied for rapid detection ofB. p um ilus for the first tmi e by incubation at 63 for only 1 h. LAMP amplification products had a ladder-like ap- pearance hen electrophoresed on an agarose ge.l The de