国境口岸13蚊种COⅠ部分基因序列分析及应用  

Sequence analysis of COⅠ partial gene among 13 mosquito species at frontier port and its potential application

作　　者：廉国胜[1] 莫秋华[1] 柯明剑[1] 汪海波[1] 田绿波[2] 杨泽[1] 林继灿[1] 苏影[1] 

机构地区：[1]珠海出入境检验检疫局国际旅行卫生保健中心,广东珠海519020 [2]四川国际旅行卫生保健中心,四川成都610000

出　　处：《热带医学杂志》2015年第10期1321-1325,共5页Journal of Tropical Medicine

基　　金：国家质检总局科技计划项目(2013IK247)

摘　　要：目的分析国境口岸13蚊种的细胞色素C氧化酶亚基Ⅰ（COⅠ）基因序列,为建立蚊种分子鉴定方法奠定基础。方法设计1对扩增COⅠ部分编码区的PCR引物,对广东、海南和云南等国境口岸采集的致倦库蚊、白纹伊蚊、中华按蚊等成蚊进行PCR扩增和序列测定,分析COⅠ核苷酸的系统进化关系。结果 13种蚊种的COⅠ基因扩增片段长度均约为400 bp,A＋T含量为69.64%~72.78%。同源性比较表明,同一蚊种间核苷酸序列同源性〉95.6%;不同蚊种间COⅠ片段核苷酸序列同源性为79.1%~93.4%。COⅠ核苷酸系统进化关系显示,从种的水平上看,白纹伊蚊、致倦库蚊、中华按蚊等上述所有蚊种均聚集成簇,即同种之间呈明显的聚集,均与形态学鉴定结果相一致。结论建立的COⅠ核苷酸鉴别技术可应用于国境口岸范围内成蚊种的区分,基本上可正确反映蚊虫的系统发育关系。这可以弥补蚊虫形态特征信息量的不足等传统分类系统的缺点,为国境口岸范围内外来的或新发现的蚊种的鉴别提供分子水平的技术依据。Objective To provide basis for future molecular identification methods of mosquito species by analyzing gene sequences of cytochrome C oxidase subunit Ⅰ（COⅠ） among 13 mosquito species at frontier port. Methods One pair of specific primer was designed to amplify part of the coding region of CO Ⅰ gene of Culex quinquefasciatus,Aedes albopictus,Anopheles sinensis,and other adult mosquitoes collected from frontier ports of Guangdong,Hainan and Yunnan. PCR products were sequenced and applied for phylogenetic analysis. Results All the amplified fragment length were approximately400 bp,with a A＋T content of 69.64%~72.78%. Homology comparison showed that the nucleotide sequence homology between the same mosquito species was over 95.6% while that between different mosquito species were 79.1%~93.4%. CO Ⅰ nucleotide sequences based phylogenetic analysis showed that all of these responding mosquito species,such as Aedes albopictus,Culex quinquefasciatus,Anopheles sinensis,etc. were gathered in their own clusters,which were consistent with traditional morphological identification. Conclusion COⅠ gene based nucleotide identification techniques can be applied to distinguish different mosquito species at frontier port,which can basically correctly reflect phylogenetic relationships between different mosquito. This technique can make up the shortcomings of inadequate mosquito morphological information in traditional classification systems. It can help to identify not only common mosquito species at frontier ports but also newly discovered or invaded exotic species.

关 键 词：蚊虫 细胞色素C氧化酶亚基Ⅰ 聚合酶链式反应 序列分析 

分 类 号：R384.1[医药卫生—医学寄生虫学]