中国血吸虫病防治杂志 ›› 2019, Vol. 31 ›› Issue (6): 628-.

• 论著 • 上一篇    下一篇

青海省多房棘球绦虫分子种系发生研究

朱文君1,韩秀敏1△*,蔡其刚2,孙艳秋1,郭亚民1   

  1. 1青海省人民医院包虫病临床研究所(西宁810007);2 青海大学畜牧兽医科学院
  • 出版日期:2020-01-17 发布日期:2020-01-18
  • 作者简介:朱文君,男,医师。研究方向:棘球蚴病预防控制 韩秀敏,女,研究员。研究方向:棘球蚴病预防控制
  • 基金资助:
    青海省卫生科技计划项目(2017-wjzx-22)

Study on the molecular phylogeny of Echinococcus multilocularis in Qinghai Province

ZHU Wen-Jun1, HAN Xiu-Min1△*, CAI Qi-Gang2, SUN Yan-Qiu1, GUO Ya-Min1   

  1. 1 Clinical Research Institute of Hydatid Disease, Qinghai Provincial People’s Hospital, Xining 810007, China; 2 Academy of Animal Science and Veterinary Medicine, Qinghai University, China
  • Online:2020-01-17 Published:2020-01-18

摘要: 目的 对青海省多房棘球绦虫分离株Cox1、Nad1基因进行扩增和测序,并构建系统进化树及分子钟,为推测其进化关系及起源提供参考依据。方法 收集2017年青海省人民医院收治的22份肝多房棘球蚴病患者术后标本,对多房棘球蚴线粒体DNA Cox1、Nad1基因进行PCR扩增和测序,并与GenBank数据库中的棘球属绦虫Cox1基因和Nad1基因序列进行比对,观察种内变异情况并构建进化树和分子钟。结果 所有多房棘球绦虫标本与GenBank数据库中检索的多房棘球绦虫亚洲株Cox1、Nad1基因序列同源性均达99%以上,共获得6种不同基因型,其中有2个分离株在GenBank数据库中未找到同源性100%的基因序列。系统进化树显示,收集的多房棘球绦虫标本与多房棘球绦虫亚洲株聚类效果明显;分子钟推测青海地区多房棘球绦虫起源于9.4万年前。结论 本研究发现青海省多房棘球绦虫存在6种不同基因型,其中首次发现2种基因型。青海地区多房棘球绦虫优势株为亚洲株,起源时间约在9.4万年前。

关键词: 多房棘球绦虫, Cox1基因, Nad1基因, 分子进化, 分子钟, 青海省

Abstract: Objective To amplify and sequence Cox1 and Nad1 genes in Echinococcus multilocularis isolates from Qinghai Province, and to create phylogenetic trees and molecular clocks, so as to provide evidence for estimating the evolutionary relationships and origins of E. multilocularis in Qinghai Province. Methods Twenty?two post?surgical specimens of patients with hepatic alveolar echinococcosis were sampled from Qinghai Provincial People’s Hospital in 2017. The Cox1 and Nad1 genes were amplified from E. multilocularis samples and sequenced. Then, the gene sequences were aligned to the Cox1 and Nad1 genes of Echinococcus species in GenBank database. The intra?species variation was observed, and the phylogenetic tree and molecular clock were created. Results All E. multilocularis samples shared more than 99% genetic homology with the sequences of Cox1 and Nad1 genes from the E. multilocularis Asian strain in the GenBank database. A total of 6 genotypes were identified, including 2 isolates that had no 100% homology with the sequences of known genes in the GenBank database. Phylogenetic tree analysis revealed remarkable clustering of the E. multilocularis samples with the E. multilocularis Asian strain, and the E. multilocularis isolates from Qinghai Province were estimated to date back to 94 000 years ago by the molecular clock. Conclusions The present study characterizes 6 E. multilocularis genotypes in Qinghai Province, including 2 novel genotypes. Asian strain is the predominant strain of E. multilocularis in Qinghai Province, and the E. multilocularis isolates from Qinghai Province date back to 94 000 years ago.

Key words: Echinococcus multilocularis, Cox1 gene, Nad1 gene, Molecular evolution, Molecular clock, Qinghai Province

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