犬猫贾第虫IGS序列特异性检测方法的建立  

DEVELOPMENT OF INTERGENIC SPACERS BASED PCR ASSAY FOR DETECTION OF GIARDIA LAMBLIA

作　　者：李雅文[1] 刘远佳[1] 郑国超 谭立娉[1] 胡伟[1] 罗琴[1] 林丽琴[1] 路鹏云[1] 李国清[1] 

机构地区：[1]华南农业大学兽医学院,广州510642

出　　处：《中国动物传染病学报》2013年第5期51-56,共6页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金资助项目(30972179;31272551)

摘　　要：应用PCR技术对广州市某宠物寄养所采集的一株D型犬源贾第虫和一株F型猫源贾第虫的核糖体IGS序列进行了扩增、克隆、测序,将测序结果与GenBank已上传的贾第虫相应序列进行比对分析,基于贾第虫IGS序列建立了具有良好特异性和敏感性的PCR检测方法,并对84份临床粪样进行了检测。结果显示,D型犬源贾第虫和F型猫源贾第虫的IGS序列长分别为1355 bp和1388 bp,贾第虫IGS序列存在多态性现象,种间差异明显,可以作为区分不同基因型的分子标记;建立的PCR方法能特异性扩增犬源贾第虫核糖体IGS序列,而犬蛔虫等对照虫体DNA均不能扩增,该方法对贾第虫DNA的最小检测量为82 fg,对84份临床粪样的检出率为3.57%,比传统镜检法高出2.38%,具有一定的临床应用价值。The intergenic spacers （IGS） were amplified in PCR from nuclear ribosomal DNAs of two Giardia lamblia strains （assemble D from dog and assemble F from cat） isolated from feces of naturally infected pets in Guangzhou. The PCR products were purified, cloned and sequenced. The resulting nucleotide sequences were compared with related sequences in the GenBank databases. The IGS sequences were 1355 bp for the canine Giardia strain and 1388 bp for the feline Giardia strain. The interspecific difference in the IGS sequence might serve as a genetic marker for the identification and differentiation of Giardia lamblia. Furthermore, a PCR detection method was developed based on IGS sequence. The established PCR assay specifically amplified IGS sequence of G. lamblia but not all control parasites such as Toxocara canis etc. The minimum detection limit of G. lamblia was 82 fg. Total 84 clinical samples were tested using this method. The positive rate of clinical samples was 3.57%, higher than 2.38% by traditional method.

关 键 词：蓝氏贾第虫 犬 核糖体基因间隔区 PCR检测 

分 类 号：S852.722[农业科学—基础兽医学]