鸡卡氏杆菌的分离与16S rRNA基因的分析鉴定  

Isolation of Gallibacterium and Identification and Analysis of 16S rRNA Gene

作　　者：田明星 林艳[1] 邹年莉[1] 李敏[1] 黄勇[1,2] 

机构地区：[1]四川农业大学动物医学院,雅安625014 [2]四川农业大学动物疫病与人类健康四川省重点实验室,雅安625014

出　　处：《中国畜牧兽医》2010年第9期57-61,共5页China Animal Husbandry & Veterinary Medicine

基　　金：教育部<长江学者和创新团队发展计划>创新团队项目(IRTO848)

摘　　要：四川雅安某种鸡场种鸡出现消瘦,腹部膨大,肝脏肿大破裂出血,腹膜和输卵管炎为主要特征的疾病。为诊断和预防此病的发生,通过对细菌分离纯化、形态学观察、生化试验、药物敏感性试验鉴定出1株致病菌,应用通用引物对细菌的16SrRNA基因进行克隆和测序,测序结果于NCBI上进行BLAST比对,选择同源性较高的巴氏杆菌科15个属的29株细菌16SrRNA序列进行比对分析,构建遗传进化树。结果表明,分离纯化的细菌能在血平板上生长,革兰氏染色阴性短杆状,能发酵大多数糖和醇类,对头孢类抗生素敏感,16S rRNA序列与卡氏杆菌属细菌的同源性最高,在92.2%～99.5%之间,特别是与鸭卡氏杆菌同源在97.5%～99.5%之间。遗传进化树分析结果表明,该菌株与卡氏杆菌属细菌属于同一大分类群,与鸭卡氏杆菌位于同一个小分支,最终诊断该鸡场为卡氏杆菌感染。An infectious disease occurred in one breeding farm in Ya＇an city of Sichuan province,mainly characterized by wasting,intumescentia abdomen,Hepatomegaly and bleeding,salpingitis,oophoritis,peritonitis,and so on.For the diagnosis and prevention of the occurrence of disease,a pathogen was identified in this paper by isolation on blood agar,morphological observation,biochemical test,antibiotic sensitive test,and 16S rRNA gene was amplified,sequenced and compared with sequences of 29 bacterias of 15 genera belonging to Pasteurellaceae,phylogenetic tree was also generated.The results showed that the bacteria was G-,short rod-shaped,and sensitive to the cephalosporin antibiotics,and homology analysis revealed that identity of 16S rRNA sequence between isolated strain and other strains of Gallibacterium was 92.2% to 99.5%,especially,identity of 16S rRNA sequence between isolated strain and other strains of Gallibacterium was 97.5% to 99.5%.Phylogenetic analysis showed that this strain and strains of Gallibacterium formed a independent clade in phylogenetic tree,especially,this strain and strains of Gallibacterium anatis fell into the same sub-branch.Finally,it was diagnosed as Gallibacterium infection in the chickens of this breeding farm.

关 键 词：卡氏杆菌 分离 鉴定 16S RRNA 同源性分析 遗传进化树 

分 类 号：S852.61[农业科学—基础兽医学]