摘要
根据胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae,App)apxIVA毒素基因序列和16SrRNA序列分别设计了一对特异性引物P1P4和一对通用引物S7S10,建立了检测App全部15个血清型的复合PCR方法。对App的15个血清型国际参考株和国内的11个App菌株进行检测,都能得到363bp和692bp的两个扩增片段。而放线杆菌等13株参考菌株只能得到692bp的扩增片段。该方法能将15个血清型的App菌株鉴定到种。检测的灵敏度达9pgDNA1300CFU。用建立的方法检测临床分离的302株可疑菌株,阳性4株,与其它鉴定方法相符。结果表明复合PCR可用于App菌株的鉴定。
A muhiplex-PCR assay was developed to identify Actinobacillus pleuropneumoniae （App）. Two pairs of polymerase chain reaction （PCR） primers were designed for the 16S rRNA and the apxIVA gene, which is specific to all serotypes of App. Two PCR products of 692bp and 363bp were obtained, from the 16S rRNA and the apxIVA gene respectively, for 27 reference A. pleuropneumoniae strains. Only the 692bp fragment was amplified for closely related strains including A. lignieresii. Using the designed primers, the method is capable of detecting A. pleuropneumoniae of as low as 1.3 × 10^3CFU or 9pg DNA. For 302 suspected isolates, this muhiplex-PCR ethod correctly identified 4 A. pleuropneumoniae strains. The result suggests the use of the muhiplex-PCR for routine identification of App.
出处
《微生物学报》
CAS
CSCD
北大核心
2005年第6期966-969,共4页
Acta Microbiologica Sinica
基金
上海市科委标准专项基金资助(02DZ05026)~~
关键词
胸膜肺炎放线杆菌
复合PCR
鉴定
ActinobaciUus pleuropneumoniae, Muhiplex-PCR, Identification
