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猪胸膜肺炎放线杆菌PCR诊断方法的建立与应用
引用本文:刁有祥,丁家波,姜世金,崔治中,禚宝山.猪胸膜肺炎放线杆菌PCR诊断方法的建立与应用[J].中国兽医学报,2006,26(4):379-381.
作者姓名:刁有祥  丁家波  姜世金  崔治中  禚宝山
作者单位:1. 山东农业大学,动物科技学院,山东,泰安,271018
2. 齐鲁动物保健品厂,山东,济南,250001
基金项目:山东省重大畜禽疫病综合防治技术项目
摘    要:根据胸膜肺炎放线杆菌ApxⅣ基因设计1对引物,扩增特异的650bp棱酸片段,建立了应用PCR检测猪胸膜肺炎放线杆菌的方法。特异性试验结果表明,12个血清型的放线杆菌参考菌株均能扩增出650bp特异性的核酸片段,而大肠杆菌、多杀性巴氏杆菌、猪肺炎支原体、伤寒沙门氏菌和支气管败血性波氏杆菌的扩增结果均为阴性。敏感性试验结果表明,PCR的最低检出限量为500个放线杆菌。利用建立的PCR检测方法对22株从山东省不同地区分离的疑似胸膜肺炎放线杆菌菌株进行检测.结果14株为阳性。对感染猪病变组织的检测结果表明,病变部位不同,胸膜肺炎放线杆菌的检出率不同,其中以扁桃体的检出率最高。

关 键 词:  传染性胸膜肺炎  胸膜肺炎放线杆菌
文章编号:1005-4545(2006)04-0379-03
收稿时间:2004-09-08
修稿时间:2004年9月8日

Identification and Detection of Actinobacillus pleuropneumoniae by PCR
DIAO You-xiang,DING Jia-bo,JIANG Shi-jin,CUI Zhi-zhong,ZHUO Bao-shan.Identification and Detection of Actinobacillus pleuropneumoniae by PCR[J].Chinese Journal of Veterinary Science,2006,26(4):379-381.
Authors:DIAO You-xiang  DING Jia-bo  JIANG Shi-jin  CUI Zhi-zhong  ZHUO Bao-shan
Institution:1. Animal Science and Technology College, Shandong Agricultural University, Taian, Shandong 271018, China ; 2. Factory of Qilu Animal Health Product, Jinan 250001, China
Abstract:The Apx IV gene, a recently discovered RTX determinant of Actinobacillus pleuropneumoniae, was shown to be , species-specific. A pair of primers was designed to specifically amplified a 650 bp fragment. The PCR result of specificity assay showed that the reference strains of the 12 serotypes of Actinobacillus pleuropneumoniae were positive, No positive results appeared in other bacterial species closely related to A. pleuropneumoniae. The sensitivity result showed that as little as 500 of Actinobacillus pleuropneumoniae could be detected by PCR. The detection results for clinical samples tissue showed that Actinobacillus pleuropneumoniae DNA could be found in trachea, nasal cavitiesor and tonsils. The high diagnostic sensitivity and specificitv of PCR will make it useful in field diagnostic work and epidemic investigation.
Keywords:PCR
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