中国寄生虫学与寄生虫病杂志 ›› 2023, Vol. 41 ›› Issue (6): 756-759.doi: 10.12140/j.issn.1000-7423.2023.06.014

• 研究简报 • 上一篇    下一篇

基于COⅠ的褐云玛瑙螺广州管圆线虫系统进化分析

高澜琳*(), 谢汉国, 林陈鑫, 江典伟, 蔡武卫, 郑丹   

  1. 福建省疾病预防控制中心,福建省人兽共患病研究重点实验室,福州 350012
  • 收稿日期:2023-04-23 修回日期:2023-07-13 出版日期:2023-12-30 发布日期:2023-12-08
  • 作者简介:高澜琳(1991-),女,硕士,主管技师,从事寄生虫遗传学、寄生虫分子生物学、寄生虫免疫学研究。E-mail:510918770@qq.com
  • 基金资助:
    福建省卫生健康科技计划(2022QNA057);福建省科技创新平台建设项目(2019Y2001)

Phylogenetic analysis of Angiostrongylus cantonensis from Achatina fulica based on COⅠ sequence

GAO Lanlin*(), XIE Hanguo, LIN Chenxin, JIANG Dianwei, CAI Wuwei, ZHENG Dan   

  1. Fujian Provincial Center for Disease Control and Prevention, Fujian Provincial Key Laboratory of Zoonosis Research, Fuzhou 350012, China
  • Received:2023-04-23 Revised:2023-07-13 Online:2023-12-30 Published:2023-12-08
  • Contact: * E-mail: 510918770@qq.com
  • Supported by:
    Fujian Provincial Health Science and Technology Plan Project(2022QNA057);Science and Technology Innovation Platform Construction Project of Fujian Province(2019Y2001)

摘要:

对福州褐云玛瑙螺感染的广州管圆线虫进行遗传进化分析,并探讨其与5大洲地区广州管圆线虫分离株间的系统进化关系。在福州市市区各大公园、市民居住地附近的垃圾场、草丛和菜地等环境,采集褐云玛瑙螺,匀浆法收集广州管圆线虫Ⅲ期幼虫并进行形态学鉴定。提取幼虫DNA,PCR扩增细胞色素C氧化酶亚基Ⅰ(COⅠ)序列,测序后在GenBank数据库中进行BLAST比对。采用MEGA软件,以血管圆线虫为外群,用邻接法构建基于广州管圆线虫COⅠ序列的系统进化树。共采集褐云玛瑙螺103只,19只感染广州管圆线虫Ⅲ期幼虫,感染率为18.4%。检获广州管圆线虫Ⅲ期幼虫1 081条。PCR扩增条带长450 bp。BLAST比对结果显示,本研究获得的COⅠ序列与福建连江广州管圆线虫(GenBank登录号:AB684364.1)的序列一致性为98.2%。系统进化树分析结果显示,本研究获得的序列与福建分离株的序列聚在一个进化分支上,再与来自北美洲(GenBank登录号:MH069731.1)、南美洲(GenBank登录号:MW390969.1)、大洋洲(GenBank登录号:KU532144.1)和欧洲(GenBank登录号:MN227185.1)的参比序列聚为一个分支。福州广州管圆线虫分离株仍属于亚洲地理株,遗传分化稳定。

关键词: 褐云玛瑙螺, 广州管圆线虫, 细胞色素C氧化酶亚基Ⅰ, 序列分析, 系统进化

Abstract:

The genetic evolution of Angiostrongylus cantonensis infected by the Achatina fulica in Fuzhou was analyzed, and the phylogenetic relationship between A. cantonensis isolates from 5 continents was discussed. The A. fulica were collected from the garbage dump, grass and vegetable fields near the residential areas of major parks in Fuzhou City. The A. cantonensis stage Ⅲ larvae were collected by homogenizing method and identified by morphology. The larval DNA was extracted, and the cytochrome C oxidase subunit Ⅰ (COⅠ) sequence was amplified by PCR. After sequencing, BLAST comparison was performed using the GenBank database. The phylogenetic tree was constructed by the neighbour-joining method using MEGA software with A. vasorum as an outgroup. A total of 103 A. fulica were collected, and 19 of them were infected with stage Ⅲ A. cantonensis larvae. The infection rate was 18.4%. A total of 1 081 A. cantonensis stage Ⅲ larvae were detected. A 450 bp band was amplified by PCR. BLAST results showed that the COⅠ sequence obtained in this study was 98.2% consistent with that of A. cantonensis (GenBank accession number: AB684364.1). Phylogenetic tree analysis showed that the sequences obtained in this study converged on an evolutionary clade with the sequences of the Fujian isolates. The isolates were further clustered with the North America isolates (GenBank accession number: MH069731.1), South America isolates (GenBank accession number: MW390969.1), Oceania isolates (GenBank accession number: KU532144.1) and Europe isolates (GenBank accession number: MN227185.1) into a branch. Therefore, the Fuzhou isolate still belongs to the Asian geographical strain, and the genetic differentiation is stable.

Key words: Achatina fulica, Angiostrongylus cantonensis, Cytochrome C oxidase subunit Ⅰ, Sequence analysis, Phylogeny

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