【摘要】 根据Gen Bank中胸膜肺炎放线杆菌(APP)荚膜多糖基因序列,设计3对引物,成功建立了检测APP 2型、3型和6型的三重PCR检测方法。该三重PCR的最低核酸检测量分别为0.25、0.5和0.25 ng/μL,对猪肺炎支原体、猪传染性萎缩性鼻炎病毒、副猪嗜血杆菌的扩增结果均为阴性。对112份自然感染病猪样品的检测结果与单一PCR检测结果完全符合。结果表明,该多重PCR方法具有很好的特异性和敏感性,可用于临床APP 2型、3型和6型的检测。更多还原

【Abstract】 According to the gene sequences of capsular polysaccharide of Actinobacillus pleuropneumoniae( APP) in Gen Bank,three pairs of specific primers were designed for amplifying the three specific fragments of serotypes 2,3 and 6,respectively. The multiplex PCR assay was successfully established,the amplified products were 500 bp,950 bp and 730 bp,respectively. The lowest nucleic acid detection was0. 25,0. 5 and 0. 25 ng / μL respectively,while the amplification results of Mycoplasma hyopneumoniae,AR and Haemophilus parasuis were negative. One hundred and twelve clinical samples were detected by the multiplex PCR,and the result was consistent with that of single PCR test. The results showed that the multiplex PCR method had good specificity and sensitivity,which could be used for clinical detection of APPserotypes 2,3 and 6.更多还原