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| 基于单碱基延伸标签反应的常见食源性致病菌基因芯片检测方法的建立 | |
| 引用本文： | 陆长勇,施春雷,张春秀,陈婧,史贤明.基于单碱基延伸标签反应的常见食源性致病菌基因芯片检测方法的建立[J].生物工程学报,2009,25(4):554-559. |
| 作者姓名： | 陆长勇 施春雷 张春秀 陈婧 史贤明 |
| 作者单位： | 1. 上海交通大学农业与生物学院食品科学系,陆伯勋食品安全研究中心,上海,200240 2. 生物芯片上海国家工程研究中心,上海,201203 3. 上海交通大学农业与生物学院食品科学系,陆伯勋食品安全研究中心,上海,200240;上海食品安全工程技术研究中心,上海,201203 |
| 基金项目： | 国家科技支撑计划(No.2006BAK02A14);;上海市重大科技专项(No.07dz19508);;国家自然科学基金项目(No.30771792)资助~~ |
| 摘 要： | 针对8种常见的食源性致病菌(金黄色葡萄球菌、副溶血弧菌、单核细胞增生李斯特菌、沙门氏菌、阪崎肠杆菌、志贺氏菌、肠出血性大肠杆菌O157:H7和空肠弯曲杆菌),建立了基于单碱基延伸标签反应原理的基因芯片检测方法。筛选和整合8种食源性致病菌基因组中的特异性序列和相应PCR引物,致病菌靶DNA片段被扩增和纯化作为单碱基延伸标签反应的模板,反应产物在DNA芯片上与探针进行杂交反应,然后通过扫描基片的荧光强度进行判断。实验结果表明,可采用基于单碱基延伸标签反应的基因芯片方法同时特异性检测8种食源性致病菌,基因组DNA多重检测灵敏度可达到0.1pg,以鼠伤寒沙门氏菌为单一检测对象的细菌纯培养物灵敏度可达到5×102CFU/mL。本方法可以快速灵敏地检测食源性致病菌,为食源性疾病的诊断和防治提供了一个有效的方法。 |
| 关 键 词： | 食源性致病菌 单碱基延伸标签 基因芯片 |
| 收稿时间： | 2008/12/23 0:00:00 |
Development of single base extension-tags microarray for the detection of food-borne pathogens |
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| Changyong Lu,Chunlei Shi,Chunxiu Zhang,Jing Chen and Xianming Shi.Development of single base extension-tags microarray for the detection of food-borne pathogens[J].Chinese Journal of Biotechnology,2009,25(4):554-559. | |
| Authors: | Changyong Lu Chunlei Shi Chunxiu Zhang Jing Chen and Xianming Shi |
| Institution: | Department of Food Science and Bor Luh Food Safety Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China;Department of Food Science and Bor Luh Food Safety Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China;National Engineering Center for Biochip at Shanghai, Shanghai 201203, China;Department of Food Science and Bor Luh Food Safety Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China;Department of Food Science and Bor Luh Food Safety Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; Shanghai Engineering Research Center of Food Safety, Shanghai 201203, China |
| Abstract: | We developed single base extension-tags (SBE-tags) microarray to detect eight common food-borne pathogens,including Staphylococcus aureus,Vibrio parahaemolyticus,Listeria monocytogenes,Salmonella,Enterobacter sakazaki,Shigella,Escherichia coli O157:H7 and Campylobacter jejuni. With specific PCR primers identified and integrated for eight food-borne pathogens,target sequences were amplified and purified as template DNA of single base extension-tags reaction. The products were hybridized to microarrays and sc... |
| Keywords: | food-borne pathogens single base extension-tags (SBE-tags) microarray |
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