| 利用双重PCR-DHPLC技术检测水稻细菌性谷枯病菌的研究 |
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| 引用本文： | 朱金国,莫瑾,朱水芳,赵文军,彭梓,刘红霞,钟文英.利用双重PCR-DHPLC技术检测水稻细菌性谷枯病菌的研究[J].植物病理学报,2010,40(5):449-455. |
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| 作者姓名： | 朱金国 莫瑾 朱水芳 赵文军 彭梓 刘红霞 钟文英 |
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| 作者单位： | 1 湖南出入境检验检疫局, 长沙 410004;2 中国检验检疫科学研究院动植物检疫研究所, 北京 100029 |
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| 基金项目： | 国家"十一·五"科技支撑计划课题，国家质检总局课题 |
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| 摘 要： | 本研究建立了一种应用双重PCR结合变性高效液相色谱技术(polymerase chain reaction-denatured high performanceliquid chromatography,PCR-DHPLC)检测水稻细菌性谷枯病菌的方法。根据水稻细菌性谷枯病菌ITS序列(internal tran-scribed spacer)和gyrB基因序列,设计两对特异性PCR检测引物,对水稻细菌性谷枯病菌株和非水稻细菌性谷枯病菌株分别进行PCR-DHPLC及双重PCR-DHPLC检测,同时进行检测灵敏度及阳性菌株的同源性分析。结果显示,PCR-DHPLC检测的特异性强,灵敏度为菌浓度4×102cfu/mL,7株水稻细菌性谷枯病菌PCR产物同源性一致。该方法能简便、灵敏、高特异性地对水稻细菌性谷枯病菌进行高通量的自动化检测。
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| 关 键 词： | 双重PCR DHPLC 水稻细菌性谷枯病菌 检测 |
Duplex PCR-DHPLC for detection of Burkholderia glumae |
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| ZHU Jin-guo,MO Jin,ZHU Shui-fang,ZHAO Wen-jun,PENG Zi,LIU Hong-xia,ZHONG Wen-ying.Duplex PCR-DHPLC for detection of Burkholderia glumae[J].Acta Phytopathologica Sinica,2010,40(5):449-455. |
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| Authors: | ZHU Jin-guo MO Jin ZHU Shui-fang ZHAO Wen-jun PENG Zi LIU Hong-xia ZHONG Wen-ying |
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| Institution: | 1 Hunan Entry-Exit Inspection and Quarantine Bureau, Changsha 410004, China;2 Animal and Plant Quarantine Institute, Chinese Academy of Inspection and Quarantine, Beijing 100029, China |
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| Abstract: | In this study, we developed a duplex PCR-DHPLC method for detecting Burkholderia glumae. It was established by using one pair of primers special to ITS (16 S-23 S rRNA interspacer region)sequence and the other pair of primers special to gyrB (β-subunit polyeptide of DNA gyrase)gene. With these two pairs of specific primers, PCR-DHPLC and duplex PCR-DHPLC methods for monitoring B. glumae could detect 4×102 cfu/mL bacterial cells. The positive PCR products of 7 strains of B. glumae showed the same sequence homology. The results provided a rapid, reliable and simple effective method for detection of B. glumae. |
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| Keywords: | DHPLC |
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