16S-23S rRNA基因间隔序列PCR及RFLP对奇异变形杆菌分离株的鉴别与系统发育分析  

Identification and phyletic evolution analysis of Proteus mirabilis strains by PCR and restriction fragment length polymorphism of 16S-23S rRNA gene intergenic spacer region

作　　者：崔国林[1] 朱瑞良[1] 左雪梅[1] 钟世勋[1] 杨世发[1] 梁漫飞[1] 孙婧[1] 刘静静[1] 

机构地区：[1]山东农业大学动物科技学院山东省动物生物技术与疫病防治重点实验室,山东泰安271018

出　　处：《中国兽医学报》2013年第3期367-370,399,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：根据临床常见致病菌16S-23SrRNA基因间隔序列(ISR)两端的16S及23SrRNA保守序列设计PCR扩增的通用引物,对9株奇异变形杆菌和6株相近菌株应用通用引物PCR扩增16S-23SrRNA ISR序列。通过PCR长度多态性比较、RFLP分析以及部分序列测序比较,分析鉴别奇异变形杆菌。结果显示,PCR长度多态性可以将奇异变形杆菌同其余菌种进行区分;RFLP分析可以将所有试验菌种进行区分;部分序列测序可以对奇异变形杆菌进行分型。由此表明,16S-23SrRNA ISR序列PCR及RFLP分析可以简单、快速、准确的鉴定奇异变形杆菌。To establish a new method to identify Proteus mirabilis strains, according to the con- served sequences of 16S and 23S which located on both ends of the clinical common pathogenic bacteria 16S-23S rRNA gene intergenic spacer region （ISR）, a pair of universal primers was de- signed. Nine Proteus rnirabilis strains and six similar bacteria strains were amplified by PCR and identified by the PCR length polymorphism comparison, restriction fragment length polymorphism （RFLP） analysis and partial sequences sequencing. The result showed that Proteus rnirabilis strains could be discriminated from other similar bacteria strains by PCR length polymorphism comparison,all of test organism could be discriminated by RFLP and Proteus rnirabilis strains could be typed by partial sequences sequencing. The result indicated that the identification method based on the 16S-23S rRNA ISR, using PCR and PCR-RFLP,is very suitable for the rapid low- cost identification and discrimination of Proteus rnirabilis strains from other phylogenetically re- lated bacteria strains.

关 键 词：奇异变形杆菌 16S-23S RRNA PCR RFLP 鉴定 

分 类 号：S852.61[农业科学—基础兽医学]