| 产毒素多杀性巴氏杆菌菌落双重PCR检测方法的建立 |
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| 引用本文： | 李伟杰,赵耘,杜昕波,康凯,陈敏.产毒素多杀性巴氏杆菌菌落双重PCR检测方法的建立[J].中国预防兽医学报,2010,32(4). |
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| 作者姓名： | 李伟杰 赵耘 杜昕波 康凯 陈敏 |
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| 作者单位： | 中国兽医药品监察所,北京,100081 |
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| 摘 要： | 为建立快速特异的PCR方法以及同时检测并区分产毒素与非产毒素多杀性巴氏杆菌,本研究根据GenBank登录的多杀性巴氏杆菌KMT1基因和toxA毒素基因序列,设计合成了2对特异引物。特异性试验表明产毒素多杀性巴氏杆菌C51-6扩增出了460bp和1854bp的2条目的片段,而不产毒素多杀性巴氏杆菌、大肠埃希菌、胸膜肺炎放线杆菌、猪链球菌、支气管败血波氏杆菌、副猪嗜血杆菌和鸡白痢沙门菌的扩增均为阴性;敏感性试验表明该PCR方法能从含450CFU的菌液中扩增出相应的目的片段。同时用豚鼠皮肤坏死试验和小鼠致死试验对该PCR方法进行了验证。
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| 关 键 词： | 产毒素多杀性巴氏杆菌 双重PCR KMT1基因 toxA基因 |
Detection of toxigenic Pasteurella multocida with a colony diplex PCR assay |
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| LI Wei-jie,ZHAO Yun,DU Xin-bo,KANG Kai,CHEN Min.Detection of toxigenic Pasteurella multocida with a colony diplex PCR assay[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(4). |
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| Authors: | LI Wei-jie ZHAO Yun DU Xin-bo KANG Kai CHEN Min |
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| Institution: | LI Wei-jie,ZHAO Yun,DU Xin-bo,KANG Kai,CHEN Min(China Institute of Veterinary Drug Control,Beijing 100081,China) |
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| Abstract: | A duplex PCR assay was developed using two sets of specific primers derived from the KMT1 gene and toxA gene of Pasteurella multocida in order to differentiate toxigenic and nontoxigenic P. multocida at the same time. The assay could specifically amplify both 460 bp and 1 854 bp DNA products from thee toxigenic P. multocida C51-6,but not from nontoxigenic P. multocida,E. coli,Actinobacillus pleuropneumoniae,Streptococcus suis,Bordetella bronchiseptica,Haemophilus parasuis and Salmonella enterica subsp. It i... |
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| Keywords: | toxigenic Pasteurella multocida diplex PCR KMT1 gene toxA gene |
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