片形吸虫Cold-SSCP鉴别方法的建立  

Development of a Cold-SSCP Technique for the Identification of Fasciola Spp

作　　者：邓国祥[1] 张汉云 陈蓓蓉[1] 李秋燕[1] 林瑞庆[1] 马妮妮[1] 朱兴全[1] 

机构地区：[1]华南农业大学兽医学院,广州510642 [2]广州市番禺区市桥城区畜牧兽医站,广州511400

出　　处：《中国畜牧兽医》2007年第2期86-88,共3页China Animal Husbandry & Veterinary Medicine

基　　金：国家杰出青年基金项目(30225033);华南农业大学校长基金(2004K024);华南农业大学大学生科技创新活动项目

摘　　要：以采自我国不同地区的片形吸虫虫体为研究对象,PCR扩增出核糖体DNA(rDNA)的第一内转录间隔区(ITS-1)序列,然后采用非同位素的单链构象多态性(Cold-SSCP)方法分析PCR产物,对不同地区片形吸虫进行分子鉴定。所有样品经Cold-SSCP分析显示2种带型。样品测序及序列分析结果表明,第1种为肝片形吸虫带型,另1种为大片形吸虫带型。本研究建立了区分大片形吸虫和肝片形吸虫的Cold-SSCP方法,可用于这2种吸虫的分子流行病学调查,从而为片形吸虫分子生物学的进一步研究奠定了基础。A non-isotopic single strand conformation polymorphism （Cold-SSCP） technique was established to differentiate fasciola hepatica from fasciola g. igantica from different geographical locations in China using the first internal transcribed spacers （ITS-1） of ribosomal DNA （rDNA） as genetic marker. The partial ITS-1 was amplified by PCR from individual fasciola and the amplicons were subjected to non-denaturing gel electrophoresis. Cold-SSCP analyses showed that fhepatica and fgigantica displayed distinct SSCP profiles, allowing their equivocal differentiation from each other. Sequencing analyses supported the results of Cold-SSCP. The establishment of the Cold-SSCP approach for the identification and differentiation of fhepatica and fgigantica has important implications for further studies of fasciola spp.

关 键 词：片形吸虫 RDNA ITS-1 Cold-SSCP 

分 类 号：S852.735[农业科学—基础兽医学]