副猪嗜血杆菌病病原的分离培养与PCR鉴定

Isolation and PCR Identification of Haemophilus parasuis

根据某猪场患病猪的临床症状和剖检变化,初步诊断为副猪嗜血杆菌引起,为确诊该病,随即取患病猪组织进行实验室检查.结果从患病猪的胸水、腹水中分离到2株革兰氏阴性细小杆菌,其生化试验为:接触酶阳性,氧化酶、吲哚和脲酶阴性;发酵果糖、麦芽糖、葡萄糖和蔗糖;不分解阿拉伯糖、鼠李糖、木糖、甘露醇、乳糖.生长需要V因子.用副猪嗜血杆菌的16S rRNA的特异性PCR引物,通过PCR技术分别从该分离菌株扩增出828bp的特异基因片段,表明该分离菌株为副猪嗜血杆菌.为疫苗的研制提供了试验依据.

According to clinical symptoms and cut exemination changes, the sick pigs would have caused by Haemop h l us p arasuis . In order to isolate and identify the bacteria, the tissues of the sick pigs were sent to the laboratory. 2 small microaerophilic Gram-negative rod bacteriums were isolated from the affected animal tissues like pleural fluid and ascites. The biochemical test： catalase was positive and oxidase, urease, indol were negative for H. parasuis; fructose, maltose, glucose and sucrose were fermentable and arabinose, rhamnose, xylose, mannitol, lactose were not fermentable. NAD were needed during culturation. 828bp specific PCR fragment of H. parasuis 16S rDNA were obtained. The results suggested that the isolate was H. parasuis. It would be provided experimental basis for the research and development of vaccines