| 猪传染性胸膜肺炎放线杆菌PCR诊断方法的建立与应用 |
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| 引用本文： | 张建新,朱岩昆,马金友,赵杰,余燕,韩科芳.猪传染性胸膜肺炎放线杆菌PCR诊断方法的建立与应用[J].中国畜牧兽医,2013,40(4):69-72. |
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| 作者姓名： | 张建新 朱岩昆 马金友 赵杰 余燕 韩科芳 |
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| 作者单位： | 1. 河南师范大学水产学院,河南新乡 453003;2. 河南科技学院动物科学学院,河南新乡 453003 |
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| 基金项目： | 河南省基础前沿与技术项目，河南省科技攻关项目 |
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| 摘 要： | 根据GenBank登录的猪传染性胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae,APP)血清1~14型的表面抗原蛋白D15和16S rRNA基因序列,设计2对特异性引物,分别扩增出637、431 bp两条核苷酸片段,建立了APP的多重PCR检测方法。特异性结果表明,血清1、3、5a、8、10型能扩增出两条与预期大小的片段,而扩增的大肠杆菌、巴氏杆菌、沙门氏菌、链球菌、葡萄球菌均成阴性。敏感性试验结果表明,最低检出浓度为50/μL。对临床分离的5株疑似APP菌株进行PCR,结果表明,其中4株为阳性,1株为阴性。提示,该方法的特异性和敏感性均良好。
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| 关 键 词： | 猪传染性胸膜肺炎 D15基因 16S rRNA 聚合酶链式反应(PCR) |
Establishment and Application of PCR Method for Diagnosing Actinobacillus pleuropneumoniae |
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| ZHANG Jian-xin
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ZHU Yan-kun
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MA Jin-you
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ZHAO Jie
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YU Yan
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HAN Ke-fang.Establishment and Application of PCR Method for Diagnosing Actinobacillus pleuropneumoniae[J].China Animal Husbandry & Veterinary Medicine,2013,40(4):69-72. |
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| Authors: | ZHANG Jian-xin ZHU Yan-kun MA Jin-you ZHAO Jie YU Yan HAN Ke-fang |
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| Institution: | 1. College of Fisheries,Henan Normal University,Xinxiang 453003,China;2. College of Animal Science,Henan Institute of Science and Technology,Xinxiang 453003,China |
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| Abstract: | According to the APP included in GenBank serum surface antigen of D15 type 1 to 14 and 16S rRNA gene sequences were designed two pairs of primers, namely amplified two nucleotide fragments of 637, 431 bp, and built PCR detection method of APP. The results indicated that type-specific serum 1, 3, 5a, 8, 10 could amplify two expected fragments and Escherichia coli, Pasteurella, Salmonella, Streptococci and Staphylococci, and they were negative. Sensitivity checked out test results showed that the lowest concentration was 50/μL. 5 strains of clinical isolates strain suspected APP by PCR ,the results showed that in four of them were positive, one was negative. The specificity of PCR detection method of the experimental establishment was strong specificity, high sensitivity. |
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| Keywords: | porcine contagious pleuropneumonia D15 gene 16S rRNA polymerase chain reaction(PCR) |
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