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Position: Home > Articles > Duplex PCR-DHPLC for detection of Burkholderia glumae Acta Phytopathologica Sinica 2010,40 (5) 449-455

利用双重PCR-DHPLC技术检测水稻细菌性谷枯病菌的研究

作  者:
朱金国;莫瑾;朱水芳;赵文军;彭梓;刘红霞;钟文英
单  位:
湖南出入境检验俭疫局;中国检验检疫科学研究院动植物检疫研究所
关键词:
双重PCR;DHPLC;水稻细菌性谷枯病菌;检测
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。
译  名:
Duplex PCR-DHPLC for detection of Burkholderia glumae
作  者:
ZHU Jin-guo1,MO Jin1,ZHU Shui-fang2,ZHAO Wen-jun2,PENG Zi1,LIU Hong-xia1,ZHONG Wen-ying1 (1Hunan Entry-Exit Inspection and Quarantine Bureau,Changsha 410004,China;2Animal and Plant Quarantine Institute,Chinese Academy of Inspection and Quarantine,Beijing 100029,China)
关键词:
duplex PCR;DHPLC;Burkholderia glumae;detection
摘  要:
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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