| 实时荧光定量PCR方法检测大肠杆菌O157:H7 |
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| 引用本文： | 陈思,黄昆仑,许文涛,李媛,罗云波.实时荧光定量PCR方法检测大肠杆菌O157:H7[J].农业生物技术学报,2006,14(5):779-782. |
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| 作者姓名： | 陈思 黄昆仑 许文涛 李媛 罗云波 |
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| 作者单位： | 中国农业大学食品学院,北京,100083 |
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| 基金项目： | 国家食品安全关键技术"十五重大专项" |
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| 摘 要： | 针对大肠杆菌(Escherichia coli )O157:H7的致病基因eae设计了特异性引物，并用荧光染料SYBR GreenⅠ 进行了实时定量聚合酶链式反应（Real-time PCR）。熔解曲线显示产物特异性较强，无非目的条带和二聚体产生。并对反应程序进行了优化，确定了最佳的反应程序，最终在合适的模板浓度内得出了标准曲线。结果显示Real-time PCR 比普通PCR灵敏1 000倍。
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| 关 键 词： | 大肠杆菌O157:H7 SYBR GreenⅠ 实时荧光定量 real-time PCR |
| 文章编号： | 1006-1304（2006）05-0779-04 |
| 收稿时间： | 2005-4-29 |
| 修稿时间： | 2005-05-23 |
Real-time Quantitative PCR Detection of Escherichia coli O157:H7 |
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| CHEN Si,HUANG Kun-lun,XU Wen-tao,LI Yuan,LUO Yun-bo.Real-time Quantitative PCR Detection of Escherichia coli O157:H7[J].Journal of Agricultural Biotechnology,2006,14(5):779-782. |
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| Authors: | CHEN Si HUANG Kun-lun XU Wen-tao LI Yuan LUO Yun-bo |
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| Institution: | College of Food Science, China Agricultural University, Beijing 100083, China |
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| Abstract: | A rapid and accurate real-time quantitative polymerase chain reaction (Real-time PCR) method with SYBR GreenⅠfor detecting Escherichia coli O157:H7 was established. A pair of amplify primers were designed to amplify eae gene. The dissociation curves showed that the amplified product was very specific. The optimal reaction program and standard curve were founded. The result indicated that Real-time PCR was sensitive 1000 times more than ordinarily PCR. |
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| Keywords: | real-time PCR |
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