【摘要】 采用胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae,App)外膜脂蛋白OmlA基因序列合成了1对特异性引物,建立了用于检测App的PCR方法。结果表明:经过PCR扩增,3株App分离株均能扩增出长约950 bp的预期DNA片段,而大肠埃希氏杆菌(Escherichia coli)、沙门氏菌(Salmonellasp.)、支气管败血波氏杆菌(Bordetella bronchiseptica)、多杀性巴氏杆菌(Pasteurella multocida)、葡萄球菌(Staphylococcussp.)、绿脓杆菌(Pseudomonas aeruginosa)、变形杆菌(Proteussp.)和链球菌(Streptococcussp.)等与猪病有关的8种病原菌均为阴性;App DNA最低检出浓度可达55.0 ng/mL,且试验结果与传统的生化反应鉴定结果完全一致,符合率达100%,证明此方法特异性和敏感性强,可用于猪传染性胸膜肺炎的快速诊断。更多还原

【Abstract】 A pair of primer was designed from an OmlA gene of Actinobacillus pleuropneumoniae and selected to be the basis for the development of a specific PCR assay.The results showed that all of 3 field isolates of A.pleuropneumoniae reacted in the PCR by the amplification of a 950 bp fragment.No reaction was observed with other bacterial species related to pig diseases,e.g.Escherichia coli,Salmonella sp.,Bordetella bronchiseptica,Pasteurella multocida,Staphylococcus sp.,Pseudomonas aeruginosa,Proteus sp.and Streptococcus sp..The lowest detection limit of DNA of A.pleuropneumoniae was 55.0 ng/mL,and the result of PCR assay was entirely accorded with the result of traditional biochemical method.It was concluded that the PCR assay developed was highly specific and sensitive,and was a reliable and quick diagnostic assay for demonstration of A.pleuropneumoniae.更多还原