鼻气管鸟杆菌PCR诊断方法的建立与应用

PCR method for Ornithobacterium rhinotracheale detection

根据GenBank已发表的鼻气管鸟杆菌(Ornithobacterium rhinotracheale,ORT)16S rRNA基因序列,设计1对可扩增671 bp目的片段的引物,建立了检测ORT的PCR方法。该方法能在ORT参考菌株中扩增到特异性片段,而鸡大肠杆菌、副鸡嗜血杆菌、鸡白痢沙门氏菌、禽多杀性巴氏杆菌的扩增结果均为阴性;敏感性试验表明该方法最低检出限量为90 pg DNA。表明所建立的PCR方法具有敏感性高、特异性强的特点。利用建立的方法对分离菌株进行检测,2株为阳性。

Based on the 16S rRNA sequence in GenBank for Ornithobacterium rhinotracheale （ORT）, a pair of primers was selected for the amplification of the 671 bp fragment to establish a PCR method for ORT detection. This method could amplify the specific fragment in the reference strains, but not in chicken E. coli, Haemophilus paragallinarum, Salmonella Pullorum and Avian pasteurella multocida, of ORT. A sensitivity test indicated that the assay＇s lowest detection limit was 90 pg DNA. The PCR methodology appeared to be highly sensitive and specific for the purpose. Positive results were obtained on two isolated strains by using this newly developed PCR method.