| 复合PCR鉴定胸膜肺炎放线杆菌和多杀性巴氏杆菌方法的建立及初步应用 |
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| 引用本文： | 贺英,赵萍,储岳峰,高鹏程,张建军,逯忠新.复合PCR鉴定胸膜肺炎放线杆菌和多杀性巴氏杆菌方法的建立及初步应用[J].江西农业大学学报,2010,32(1). |
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| 作者姓名： | 贺英 赵萍 储岳峰 高鹏程 张建军 逯忠新 |
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| 作者单位： | 中国农业科学院,兰州兽医研究所/家畜疫病病原生物学国家重点实验室/农业部草食动物疫病重点开放实验室/农业部畜禽病毒学重点开放实验室,甘肃,兰州,730046 |
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| 基金项目： | 国家科技支撑计划项目(2006BAD06A12);;国家科技支撑计划项目(2006BAD06A01);;国家科技基础性工作专项(2008FY210200) |
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| 摘 要： | 在已经建立的胸膜肺炎放线杆菌、多杀性巴氏杆菌的单项PCR诊断方法的基础上,通过对扩增条件和引物的筛选,成功地建立胸膜肺炎放线杆菌(App)、多杀性巴氏杆菌(PM)复合PCR诊断方法,利用1次PCR反应,即可同时扩增App的342bp和PM的457bp的特异性片段。检测灵敏度分别达9.7pgDNA/1.4×103OCFU和16pgDNA/2.3×103OCFU,用建立的方法检测临床分离的23株可疑菌株,App阳性8株,PM阳性2株,与其它鉴定方法检测结果相符,表明该方法的建立能够对这2种疾病进行临床鉴别诊断。
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| 关 键 词： | 胸膜肺炎放线杆菌 多杀性巴氏杆菌 复合PCR 应用 |
Development and Application of Multiplex-PCR for Identification of Actinbacillus pleuropneumoniae and Haemophilus parasuis |
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| HE Ying,ZHAO Ping,CHU Yue-feng,GAO Peng-cheng,ZHANG Jian-jun,LU Zhong-xin.Development and Application of Multiplex-PCR for Identification of Actinbacillus pleuropneumoniae and Haemophilus parasuis[J].Acta Agriculturae Universitis Jiangxiensis,2010,32(1). |
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| Authors: | HE Ying ZHAO Ping CHU Yue-feng GAO Peng-cheng ZHANG Jian-jun LU Zhong-xin |
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| Institution: | Lanzhou Veterinary Research Institute/a>;Chinese Academy of Agricultural Sciences/State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture Key La-boratory of Animal Virology of Ministry of Agriculture/a>;Lanzhou 730046/a>;China |
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| Abstract: | The multiplex PCR detecting App and PM were developed based on published single PCR for these pathogens.The PCR reagents,annealing temperature and the PCR procedure were optimized and the primers were screened.A 342 bp fragment of App,and a 457 bp of PM could be simultaneously amplified in one reaction.App was as low as 1.4×10~3 CFU or 9pg DNA and PM was as low as 2.3×10~3 CFU or 16pg DNA.For 23 suspected isolates,this multiplex-PCR method correctly identified 4 App and 2 PM.The result suggests the usefulness of the muhiplex PCR for routine identification of App and PM. |
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| Keywords: | Actinbacillus pleuropneumoniae Pasteurella multocida multiplex-PCR application |
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