【作者】 马晓平； 曹三杰； 文心田； 肖国生； 杨利； 陈华美； 张雪峰；

【Author】 MA Xiao-ping,CAO San-jie,WEN Xin-tian,XIAO Guo-sheng,YANG Li,CHEN Hua-mei,ZHANG Xue-feng (Laboratory of Animal Infectious Disease and Microarray,Key Laboratory of Animal Disease and Human Health of Sichuan Province,College of Veterinary Medicine, Sichuan Agricultural University,Ya’an,Sichuan 625014,China)

【机构】 四川农业大学动物医学院动物传染病与基因芯片实验室四川省动物疫病与人类健康重点实验室；

【摘要】 本试验根据已经发表的猪传染性胸膜肺炎放线杆菌dsbE-like基因的序列,设计了一对可扩增374bp目的片段的特异性引物.经PCR扩增,APP1-4和6-10型标准菌株均能扩出大约374bp的片段,而大肠杆菌(K88)、巴氏杆菌(5:A)和链球菌(Ⅱ)等的扩增结果均为阴性.将此方法用来检测分离菌株MC、ME和MH,结果表明该方法能快速检测出临床分离株(APP).灵敏性试验表明:本方法对APP菌液最低检出浓度为71CFU/mL.结果表明:建立的PCR检测APP的方法能快速、灵敏地检测出APP标准株和临床分离株,本方法可用于APP的流行病学调查和临床诊断.更多还原

【Abstract】 According to the sequences of porcine Actinobacillus pleuropneumoniadsbE-like gene reported in Genebank,a pair of primer to amplify fragment of 374bp were designed and synthesized.From reference strains APP1～4 and 6～10,fragments as expected of 374bp were successfully amplified while failed to gain the same result from E.coli(K88),Pasterurella(5:A)and Streptococcus(Ⅱ).Further experiments on isolates MC,ME and MH demonstrated that this method could be used to quickly detect A.pleuropneumoniae from clinical samples.Sensitivity test demonstrated that the lowest concentration of APP which could be detected was 71CFU/mL.In other words,such kind of PCR method for the detection of A.pleuropneumoniae could be applied for epidemiological investigation or clinical diagnosis of APP.更多还原