钩棘单睾吸虫和扇棘单睾吸虫ITS2序列测定及其种系发育分析  

ITS2 Sequencing and Phylogenetic Analysis of Haplorchis pumilio and H.taichui

作　　者：梅雪芳[1] 李树清[2] 胡长红 黄腾飞 陈志飞[2] 黄维义[1] 

机构地区：[1]广西大学动物科技学院,南宁530005 [2]上海出入境检验检疫局,上海200135 [3]罗氏诊断产品(上海)有限公司,上海200335

出　　处：《中国畜牧兽医》2015年第8期1943-1949,共7页China Animal Husbandry & Veterinary Medicine

基　　金：上海出入境检验检疫局科研项目(HK002-2014);国家质检总局科技专项(2010IK013)

摘　　要：本试验将从犬猫肠道中分离的钩棘单睾吸虫和扇棘单睾吸虫用盐酸卡红染色后观察其形态学特征;PCR扩增其ITS2基因序列并测序,并将测序结果与GenBank同属异形科的多棘单睾吸虫和横川后殖吸虫序列进行比对;应用Mega 6.05软件采用邻位相连法构建种系发育树进行分析。盐酸卡红染色后显示,钩棘单睾吸虫和扇棘单睾吸虫各形态学特征符合经典的分类学描述。PCR测序后获得钩棘单睾吸虫和扇棘单睾吸虫ITS2序列长度分别为295和297bp,G＋C含量分别为49.5%（146/295）和54.2%（161/297）。提交至GenBank后获得登录号分别为钩棘单睾吸虫KJ137221-KJ137223,KP165437-KP165439;扇棘单睾吸虫KP165440。序列比对结果显示,钩棘单睾吸虫和扇棘单睾吸虫种内差异性为0,4种异形吸虫种间差异为15.2%-28.9%。基于ITS2序列建立的种系发育树显示钩棘单睾吸虫和扇棘单睾吸虫构成自展值为78%的拓扑分支。The morphological features of Haplorchis pumilio （H. pumilio） and H. taichui were observed with hydrochloric acid carmine staining; The ITS2 sequences were amplified by PCR, analyzed by sequencing, and compared with sequences of H. yokogawai and Metagonimus yokogawai of Heterophyidae trematodes downloaded from GenBank And the phylogenetic tree was con- structed by Mega 6.05. The morphological characteristics of H. pumilio and H. taichui from dog and cat were conformed to the classical taxonomic description. The ITS2 sequences of H. pumilio and H. taichui were 295 and 297 bp,and the percentages of G＋C content of ITS2 were 49.5% （146/295） and 54.2% （161/297）,respectively. The sequences were submitted to GenBank and the accession numbers of H. pumilio were KJ137221-KJ137223 and KP165437-KP165439, the accession number of H. pumilio was KP165440. The sequence alignment results showed that H. purnilio and H. taichui intraspecific difference was 0,interspecific differernce with four heterophyes was 15.2% to 28.9%. The phylogenetic tree based on ITS2 showed that H. pumilio and H. taichui were at the same clade with the bootstrap value of 99%.

关 键 词：钩棘单睾吸虫 扇棘单睾吸虫 ITS2基因 种系发育分析 

分 类 号：Q78[生物学—分子生物学]