Liu W, Liu G H, Li F, He D S, Wang T, Sheng X F, Zeng D L, Yang F F, Liu Y
College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan Province 410128, PR China.
J Helminthol. 2012 Sep;86(3):271-5. doi: 10.1017/S0022149X1100037X. Epub 2011 Jul 20.
Sequence variability in three mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 3 (cox3), NADH dehydrogenase subunits 1 and 4 (nad1 and nad4) in Spirometra erinaceieuropaei spargana from different geographical regions in China was examined. A portion of each of the cox3 (pcox3), nad1 (pnad1) and nad4 genes (pnad4) were amplified separately from individual S. erinaceieuropaei spargana by polymerase chain reaction (PCR). Representative amplicons were subjected to sequencing in order to estimate sequence variability. The sequences of pcox3, pnad1 and pnad4 were 541, 607 and 847 bp in length, respectively. The A+T contents of the sequences were 68.39-68.76% (pcox3), 63.76-64.91% (pnad1) and 67.18-67.77% (pnad4), respectively, while the intra-specific sequence variations within each of the S. erinaceieuropaei spargana were 0-1.5% for pcox3, 0-2.8% for pnad1 and 0-2.7% for pnad4. Phylogenetic analysis using neighbour joining (NJ), maximum likelihood (ML) and maximum parsimony (MP) methods, indicated that all the spargana isolates in Hunan Province represented S. erinaceieuropaei. These findings demonstrated clearly the usefulness of the three mtDNA sequences for population genetics studies of S. erinaceieuropaei spargana of human and animal health significance.
对来自中国不同地理区域的曼氏迭宫绦虫裂头蚴三个线粒体DNA(mtDNA)区域,即细胞色素c氧化酶亚基3(cox3)、NADH脱氢酶亚基1和4(nad1和nad4)的序列变异性进行了检测。通过聚合酶链反应(PCR)分别从单个曼氏迭宫绦虫裂头蚴中扩增出cox3(pcox3)、nad1(pnad1)和nad4基因(pnad4)的一部分。对代表性扩增子进行测序以估计序列变异性。pcox3、pnad1和pnad4的序列长度分别为541、607和847 bp。这些序列的A+T含量分别为68.39 - 68.76%(pcox3)、63.76 - 64.91%(pnad1)和67.18 - 67.77%(pnad4),而曼氏迭宫绦虫裂头蚴个体内的种内序列变异在pcox3中为0 - 1.5%,在pnad1中为0 - 2.8%,在pnad4中为0 - 2.7%。使用邻接法(NJ)、最大似然法(ML)和最大简约法(MP)进行的系统发育分析表明,湖南省的所有裂头蚴分离株均代表曼氏迭宫绦虫。这些发现清楚地证明了这三个mtDNA序列对于对人类和动物健康具有重要意义的曼氏迭宫绦虫裂头蚴群体遗传学研究的有用性。