鸡奇异变形杆菌的分离鉴定和16S rRNA基因序列测定与系统进化分析  

Isolation,identification of chicken Proteus mirabilis and phylogenetic analysis of 16S rRNA gene sequence

作　　者：朱明华[1] 朱瑞良[1] 马荣德[1] 孙寅剑[1] 郭文龙[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医学报》2011年第6期804-808,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972183)

摘　　要：从山东泰安一鸡场发病雏鸡眼中分离到1株致病菌(编号为QY),通过细菌形态学等常规鉴定符合奇异变形杆菌(Proteus mirabilis)特性。用奇异变形杆菌阳性血清诊断结果呈阳性,人工感染证明该菌株是造成该鸡场雏鸡大批发病死亡的致病菌。药敏试验结果显示对头孢类、恩诺沙星等高度敏感,而对青霉素和复合磺胺等不敏感。以细菌16SrRNA基因通用引物进行PCR扩增,得到QY的16SrRNA基因序列,长约1 453bp(GenBank,登录号为GU477712)。将该序列与GenBank中序列进行Blast比对,发现与其匹配度最高的均是奇异变形杆菌各株系的16SrRNA序列,均高达98%以上。运用DNAStar软件与其中10株奇异变形杆菌分离株构建系统进化树,结果表明,分离株(QY菌株)与10个代表菌株的同源性均为98.9%～99.9%,其中与AB272366同源性最高为99.9%。从分子水平证明该菌是奇异变形杆菌并分析了其遗传进化规律,为鸡奇异变形杆菌的鉴定及其引起的疾病的诊断与治疗提供了参考。One strain disease-causing bacteria marked QY was isolated from the infected chicken in Tai′an Shandong province,and identified as P.mirabilis by traditional method of isolation and identification.P.mirabilis serological diagnostic reaction was positive of infected animal experiments proved that QY was the major pathogen that caused the chicken a large number of sick and death in the local area.Antibiotic susceptibility test showed that it was highly sensitive to Cephalosporin and Enrofloxacin while resistent to Penicillin and Sulfa.The 16S rRNA gene of 1 453 bp was amplified from QY by PCR,which was deposited in GenBank（No.GU477712）.Blast analysis showed that the gene shared more than 98.9% homology with the sequences of Proteus mirabilis.then built phylogenetic tree with 10 strains of P.mirabilis using DNAStar.Showing that the homology were 98.9%-99.9%,and the homology with AB272366 was up to 99.9%,the QY was confirmed as P.mirabilis from the molecular level.it provided reference for the diagnosis and treatment of infected chicken and the identification of P.mirabilis.

关 键 词：鸡奇异变形杆菌 16S RRNA 系统进化树 同源性 

分 类 号：S852.61[农业科学—基础兽医学]