DNA条形码技术在蚤类鉴定中的应用

Application of DNA barcoding technique in the molecular identification of Siphonaptera

目的以中国-哈萨克斯坦(中哈)边境阿拉山口地区常见蚤种为研究对象,使用线粒体细胞色素C氧化酶亚基Ⅰ(COⅠ)基因对其进行分子生物学鉴定。方法 2017年5月在中哈边境阿拉山口地区采集鼠体表寄生蚤,形态学鉴定完毕后提取DNA,使用PCR方法扩增线粒体COⅠ并测序。将测序结果利用BLAST分析,使用MAGE 6.0软件构建分子进化树。结果长吻角头蚤和猫栉首蚤指名亚种形态学与分子生物学鉴定100%相符,并首次在国际上报道臀突客蚤和秃病蚤指名亚种的线粒体COⅠ基因序列。4种蚤类分子生物学鉴定与形态学鉴定相吻合,中哈边境地区的4种蚤类与国外的蚤类有着高度相似性。结论 DNA条形码技术能够对蚤类进行有效的种类鉴定,为口岸快速准确地识别宿主和外来医学媒介生物的防控提供科学依据。

Objective To conduct molecular identification of the common Siphonaptera species from the Alataw pass of China-Kazakhstan border based on the mitochondrial cytochrome C oxidase subunitⅠ(COⅠ)gene.Methods In May 2017,parasitic Siphonaptera on the body surface of rodents was collected from the Alataw pass of China-Kazakhstan border.DNA was extracted after morphological identification.The mitochondrial COⅠgene was amplified by PCR and sequenced.The sequencing results were analyzed by BLAST,and the phylogenetic tree was constructed using MAGE 6.0 software.Results The morphological and molecular biological identification showed 100%consistency between Echidnophaga oschanini and Ctenocephalides felis felis.Meanwhile,the mitochondrial COⅠsequences of Xenopsylla minax and Nosopsyllus laeviceps laeviceps were reported for the first time in the world.Molecular biological identification and morphological identification of the four species of Siphonaptera were in good agreement.The four species of Siphonaptera in China-Kazakhstan border were highly similar to Siphonaptera species in other countries.Conclusion DNA barcoding technique can identify the species of Siphonaptera effectively and provide a scientific basis for rapid and accurate identification of host and control of foreign medical vectors at ports.