东北地区华支睾吸虫ITS1基因序列分析

摘要

To examine the DNA polymorphism in ITS1 of Clonorchis sinensis from different areas, C.sinensis isolates were collected from Binxian, Daqing, Hailun, Shuangcheng, Tailai, Tongjiang and Changchun areas, respectively.The complete ITS1 sequence of these isolates were sequenced and thc ITS1 sequence were 661 bp in all the isolates with identity of 99.4％ to 100％.The neighbor-joining tree was established by comparing with thc ITS1 sequences of Opisthorchis viverrini, Opisthorchis, felineus, Metorchis orientalis, C.sinensis (Guangxi, Shenyang and Korea isolates) in GenBank, respectively.The phylogenetic tree by NJ algorithm revealed that Guangxi isolate was on a single clade and relatively far from others.The Korea isolate was closely related to thc isolates from Northeast China.The isolates of Hailun and Tailai, Daqing and Shenyang, Changchun and Tongiiang, Shuangcheng and Binxian were clustered in the same clade, respectively.This study showed that ITS1 could be applied as genetic marker in identifying intergeneric genetic relation and classifying interspecific genetic relation of C.sinensis.%为研究我国东北地区华支睾吸虫(Clonorchis sinensis)ITS1基因的变异情况及多态性,本研究以采自宾县、大庆、海伦、双城、泰来、同江和长春7个地区的华支睾吸虫为研究对象,PCR扩增ITS1基因,并与GenBank登录的麝猫后睾吸虫、猫后睾属吸虫、东方次睾吸虫、广西株华支睾吸虫、沈阳株华支睾吸虫和韩国株华支睾吸虫的ITS1基因进行比对分析.结果表明,各地区华支睾吸虫样品ITS1基因大小均为661bp,同源性在99.4%～100%之间.构建的系统发生树显示,分支情况与地区距离呈正相关,广西株与其它地区华支睾吸虫所属分支相隔较远,韩国株与东北地区各株相隔较近,海伦株与泰来株,大庆株与沈阳株,长春株与同江株,双城株与宾县株分别位于同一分支.结果显示,ITS1片段除了可作为遗传标记用以鉴定华支睾吸虫科内属间遗传关系,还可以区分属下种间的遗传关系.