摘要
为鉴定我国青藏高原分离的田鼠源绦虫囊尾蚴的种类及其遗传进化特征,本研究通过对该囊尾蚴进行形态学观察,采用绦虫cox1基因通用引物,利用PCR扩增并测序,初步鉴定该囊尾蚴。形态学观察结果显示,该囊尾蚴为小型虫体,虫体背部和腹部稍扁平,长度约为200.0 mm,虫体横截面直径约为2.5 mm;虫体分为3部分:前端部分直径通常略小,中间部分表面为皱褶区域,约占整个虫体的一半,后端部分细长,相对光滑,有平行的横向凹陷;PCR结果显示,在约440 bp处出现与预期目的片段大小相符的条带,测序结果显示,该基因序列与多刺带绦虫(T.polyacantha)cox1基因序列(EU544581)的同源性高达97.0%。初步表明该囊尾蚴属于多刺带绦虫囊尾蚴。采用多刺带绦虫线粒体cox1、nad1和12S rRNA特异性基因引物经PCR进一步鉴定并测序。将3个基因测序结果与GenBank登录的多刺带绦虫的相应基因序列比对,分析它们之间的同源性,并构建这3个基因的进化树,分析它们之间的遗传进化关系。PCR结果显示,分别扩增到约390 bp(cox1)、480 bp(nad1)和240 bp(12S rRNA)的目的条带。同源性分析结果显示,3个基因序列与GenBank登录的多刺带绦虫相应基因序列的同源性最高分别达99.4%、98.7%和100.0%。进化树结果显示,该虫株的3个基因均与不同宿主的多刺带绦虫处于同一分支。本研究在国内首次鉴定到青藏高原田鼠源多刺带绦虫囊尾蚴,并分析其线粒体基因的同源性及遗传演化关系,为多刺带绦虫的分类、鉴定和种群遗传进化分析及分子流行病学调查等奠定了研究基础。
In order to identify the species and genetic evolution characteristics of Taenia polyacantha cysticercus isolated from vole in the Qinghai-Tibet Plateau,this study preliminarily identified the cysticercus by observing its morphological characteristics,and using universal primers for the tapeworm cox1 gene amplification.Morphological characteristics observation results showed that the cysticercus was a small worm,with a slightly flat back and abdomen,a length of about 200.0mm,and a cross-sectional diameter of about 2.5mm;the cysticercus was mostly divided into three parts:the front part was usually slightly smaller in diamater,the surface of the middle part was a wrinkled area,accounting for about half of the entire length of the insect body,and the rear end part was slender,relatively smooth,and had parallel lateral depressions.The PCR results showed that a band consistent with the size of the expected target fragment appeared at about 440bp.The sequencing results showed that the homology of this gene sequence with the cox1 gene(EU544581)of the T.polyacantha was as high as 97.0%.Preliminary results indicated that the cysticerci belonged to T.polyacantha cysticerci.The specific primers to mitochondrial cox1,nad1 and 12S rRNA gene of T.poly-acantha were used for PCR amplification.The sequencing results of the three genes were compared with the corresponding gene sequences of T.polyacantha in GenBank,and the homology between them was analyzed.The evolutionary trees of these three genes were constructed to analyze the genetic evolutionary relationship.The PCR results showed that the target bands of approximately 390bp(cox1),480bp(nad1)and 240bp(12S rRNA)were amplified respectively.The results of homology analysis showed that the homology between the three gene sequences and the corresponding gene sequences of T.polyacantha registered in the GenBank database was up to 99.4%,98.7%and 100.0%,respectively.The evolutionary tree results showed that the three genes of this strain were in the same branch as T.polyacantha from different hosts.This study was the first in China to identify T.polyacantha cysticercus from vole and analyze its mitochondrial gene homology and genetic evolution relationship.This study provides a basis for the classification,identification,population genetic evolution analysis and molecular epidemiological investigation of the T.polyacantha.
作者
张学勇
简莹娜
朵红
郭志宏
马霄
张青
赵存哲
郭帅
马万里
李志
付永
ZHANG Xue-yong;JIAN Ying-na;DUO Hong;GUO Zhi-hong;MA Xiao;ZHANG Qing;ZHAO Cun-zhe;GUO Shuai;MA Wan-li;LI Zhi;FU Yong(The Academy of Animal and Veterinary Sciences,Qinghai University,Qinghai Provincial Key Laboratory of Pathogen Diagnosis for Animal Disease and Green Technical Research for Prevention and Control,Xining 810016,China;Qinghai Province Institute for Endemic Diseases Prevention and Control,Xining 810016,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2023年第11期1193-1198,共6页
Chinese Journal of Preventive Veterinary Medicine
基金
青海省科技厅应用基础研究项目(2021-ZJ-724)
青海省“昆仑英才·高端创新创业人才”项目。
关键词
多刺带绦虫
囊尾蚴
田鼠
鉴定
遗传进化
Taenia polyacantha
cysticercus
vole
identification
phylogenetic analysis
