鸭疫里默氏菌的分离培养和PCR鉴定

Separation and PCR Identification of Riemerella anatipestifer

对某鸭场送检的发病雏番鸭进行临床症状、病理变化观察、病原的分离和纯化,同时以外膜蛋白(OMPA)保守区设计一对特异性引物,PCR扩增,回收目的条带,进行测序与序列分析。结果表明,该菌为革兰氏阴性、无芽孢小杆菌,在TSA培养基上形成光滑圆形透明菌落,有荧光,不发酵糖,初步怀疑为鸭疫里默氏菌;PCR扩增,琼脂糖凝胶电泳,在600bp处扩增出了一条清晰的条带,回收目的条带,测序,并与genebank中序列比对,确诊该菌为鸭疫里默氏菌。

The clinical symptoms and pathological change were observed from the sick young Muscovy ducks that were got from the farm. Then separation and purification of pathogens were carried on. Meanwhile a pair of specificity primer was designed according to the outside membrane protein conservative district of the bacterium, and the gene was amplified by PCR. Then the purpose sequence was sequenced and analyzed. The results showed that the bacterium was gram-negative , non-spore-bearing,eubacterium minutum. And the smooth circular transparency bacterial colony was observed on the TSA culture medium. There was fluorescence on the surface of the bacterium. The bacterium can not ferment the sugar. According to those specialities,the bacterium was thought to RiemereUa anatipestifer. Then a 600 bp fragment was amplified by PCR, and the gene sequence was sequenced. Sequence alignment was done in genebank. According to that the bacterium was identified to Riemerella anatipestifer.