贝氏柯克斯体毒力相关基因芯片对其他胞内寄生菌基因组DNA检测结果分析  

Analysis of DNA of related isolates with virulence-associated microarray of Coxiella burnetii

作　　者：王世斌[1,2] 赵建忠[1] 董晓根[1] 

机构地区：[1]北京市丰台区疾病预防控制中心,北京100071 [2]军事医学科学院微生物流行病研究所,北京100071

出　　处：《热带医学杂志》2012年第9期1057-1061,共5页Journal of Tropical Medicine

基　　金：国家自然科学基金(30670101);中国博士后基金(20080441328)

摘　　要：目的用建立的贝氏柯克斯体毒力相关基因芯片研究贝氏柯克斯体与其致病性相似的专性胞内寄生菌(立克次体、埃立克体、新立克次体),以及与其密切相关的兼性胞内寄生菌(巴通体和肺炎军团菌)等病原菌的毒力基因的差异。方法贝氏柯克斯体国际标准株-九里株作为本研究参考株,从全基因序列中选取166个毒力及毒力相关基因进行PCR扩增,纯化后的产物制备基因芯片。采用双色荧光标记,待测DNA(即用于分析的各菌株基因组DNA)用Cy5标记,贝氏柯克斯体九里株参照基因组DNA用Cy3标记。芯片杂交后,采用软件GenePixPro4.1,辅助以软件Excel,进行图片处理和数据分析。结果芯片的166个基因片段中有165个存在于所有贝氏柯克斯体属的6个分离株中,另一锚定蛋白(AnkI)基因(CBU1213)仅存在于九里株及Grita株;立克次体属的5株菌株均与CBU1631和CBU1125杂交;埃立克体属的3种菌株与新立克次体属的3种菌株均含有基因CBU1125;东方属的1株菌株具有基因CBU1449,而巴通体属的菌株,军团菌和大肠杆菌与基因芯片均无杂交。结论贝氏柯克斯体种内基因组具有高度保守性,它的专性细胞内寄生菌的毒力相关基因与胞外寄生菌的毒力相关基因有非常显著差异。Objective To study the differences between the Coxiella burnetii and the the obligate intracellular parasites showing pathogenesis（the genus Rickettsia Ehrichia,Neorickettsia）,and the closely related facultative parasite（Bartonella and Legionella pneumophila）,using the virulence-associated microarray.Methods 166 virulence-related genes implicated in adhesion,invssion,intracellular trafficking,host modulation,detoxification were selected from the whole genome of Nine Mile of Coxiella burnetii and processed by PCR.The PCR products were purified and made the microarray.By methods of twofluorescence comparative hybridization the microarray was labeled,the reference DNA（DNA of Nine Mile）was labeled by Cy3,to be analysed DNA were labeled by Cy5.After the hybridization,processing image and data analysing based on the GenePix Pro4.1 and Excel software.Results The 165 genes hybridized in 6 isolates of Coxiella burnetii hybridized with the microarray with 166 virulence-related genes,the AnkI gene（CBU1213） exists only in Nine Mile or Grita;5 isolates of Rickettsia positively hybridized with CBU1631 and CBU1125;3 isolates of Ehrichia and 3 isolates of Neorickettsia all contain CBU1125;A isolate of Orientia has gene of CBU1449,The genomic DNA of Bartonella,Escherichia coli and Legionella pneumophila negatively hybridized with the 166 virulence-related genes.Conclusion The virulence-associated genes among C.burnetii strains were strictly conserved.Their virulence-associated genes between obligate intracellular parasites and extracellular parasite are difference.

关 键 词：贝氏柯克斯体 毒力基因 基因芯片 杂交 

分 类 号：R376.4[医药卫生—病原生物学]