Department of Veterinary Science, The University of Melbourne, 250 Princes Highway, Werribee, Victoria 3030, Australia.
Mol Cell Probes. 2010 Dec;24(6):346-51. doi: 10.1016/j.mcp.2010.07.005. Epub 2010 Jul 24.
We genetically classified Echinococcus granulosus from humans, cattle and camels in Libya utilizing DNA regions (designated pcox1 and pnad1) within the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase 1 (nad1) mitochondrial genes, respectively. Polymerase chain reaction (PCR)-based single-strand conformation polymorphism (SSCP) analysis of pcox1 and pnad1 amplicons derived from genomic DNA samples from individual cysts (n = 176) revealed four distinct electrophoretic profiles for each locus. Direct sequencing of selected amplicons representing each of these profiles defined four different sequence types for each locus, which were present in five different combinations (designated haplotypes A-E) amongst all 176 isolates. Phylogenetic analysis of concatenated sequence data for these five haplotypes, together with a range of well-defined reference sequences, inferred that all cyst isolates from humans (n = 55) and a small number from cattle (13% of 38) belonged to the G1-G3 complex of E. granulosus (or E. granulosus sensu stricto), whereas most (87%) cysts from cattle and all 83 of them from camels were linked to the G6-G10 complex (or Echinococcus canadensis). The present study provides a foundation for future large-scale studies of the epidemiology and ecology of E. granulosus in Libya and other African countries.
我们利用线粒体基因细胞色素 c 氧化酶亚基 1 (cox1) 和 NADH 脱氢酶 1 (nad1) 内的 DNA 区域(分别指定为 pcox1 和 pnad1),对来自利比亚的人类、牛和骆驼中的细粒棘球绦虫进行了基因分类。从个体囊肿(n=176)的基因组 DNA 样本中获得的 pcox1 和 pnad1 扩增子的聚合酶链反应(PCR)-基于单链构象多态性(SSCP)分析显示,每个基因座都有四种不同的电泳图谱。对代表每种图谱的选定扩增子进行直接测序,确定了每个基因座的四种不同序列类型,这些类型存在于所有 176 个分离株中的五种不同组合(命名为单倍型 A-E)中。对这五个单倍型的串联序列数据进行系统发育分析,以及一系列明确的参考序列,推断所有来自人类的囊肿分离株(n=55)和一小部分来自牛(38%的 13%)属于细粒棘球绦虫的 G1-G3 复合物(或细粒棘球蚴),而大多数(87%)来自牛的囊肿和来自骆驼的全部 83 个囊肿都与 G6-G10 复合物(或加拿大棘球蚴)有关。本研究为未来在利比亚和其他非洲国家进行细粒棘球蚴流行病学和生态学的大规模研究奠定了基础。