DNA条形码技术在黑瞎子岛蜱类鉴别中的应用  

Application of DNA barcoding technology in identification of ticks in Heixiazi Island

作　　者：程成 王艳梅 翟琪安 鞠文东 焦丹 徐宁 耿聪 韩茜宇 于晓维 黄洋[4] CHENG Cheng;WANG Yan-mei;ZHAI Qi-an;JU Wen-dong;JIAO Dan;XU Ning;GENG Cong;HAN Xi-yu;YU Xiao-wei;HUANG Yang(Heilongjiang International Travel Healthcare Center,Harbin,Heilongjiang 150001,China;Northeast Forestry University;Heihe Customs;Heilongjiang Red Cross Sengong General Hospital)

机构地区：[1]黑龙江国际旅行卫生保健中心,黑龙江哈尔滨150001 [2]东北林业大学 [3]黑河海关 [4]黑龙江省森工总医院

出　　处：《中国公共卫生管理》2020年第5期675-678,共4页Chinese Journal of Public Health Management

基　　金：海关总署科研计划项目科研基金(2017IK158;2018IK014)。

摘　　要：目的应用DNA条码技术对黑瞎子岛蜱样本进行鉴定,为蜱的种类鉴定提供更加准确快速的依据。方法对捕获的557只蜱样本完成形态学鉴定后,提取蜱样本基因组DNA,采用PCR方法从蜱基因组中扩增线粒体细胞色素c氧化酶亚基Ⅰ(COⅠ)基因,测序后用Mega6.0软件构建系统进化树。结果经形态学鉴定为1科3属4种,分别是森林革蜱(DS1和DS2)、全沟硬蜱(IP1和IP2)、嗜群血蜱(HC1和HC2)和日本血蜱(HJ)。编号IP1和IP2样本与GenBank中编号JQ737101.1和编号KU935457.1公布的全沟硬蜱COⅠ序列相似性为99.85%~100.00%;编号HC1和HC2样本与GenBank中编号KR108861.1公布的嗜群血蜱COⅠ序列相似性为99.84%;编号HJ样本与GenBank中编号KR108879.1公布的日本血蜱COⅠ序列相似性为99.84%;以上结果均与形态学鉴定结果一致。编号DS1和DS2样本与GenBank中编号KU594270.1公布的草原革蜱COⅠ序列相似性为99.84%,与编号MH079424.1公布的森林革蜱COⅠ序列相似性为99.22%,二者虽均符合可直接鉴定标准,但综合草原革蜱和森林革蜱的形态学特征,可以确定样本均为森林革蜱。结论在传统形态学鉴定分类的基础上,结合DNA条码技术,能准确快速鉴定蜱的种类。Objective To identify the samples of ticks in Heixiazi Island by using DNA barcoding technology, in order to provide more accurate and rapid basis for species identification of ticks. Methods After the morphological identification of 557 captured ticks, genomic DNA of tick samples was extracted, and the mitochondrial cytochrome c oxidase subunit I(COⅠ) gene was amplified from tick genome by PCR, phylogenetic tree was constructed with Mega6.0 software after sequencing. Results They were morphologically identified as one family, three genera and four species, which were Dermatocentor silvarum(DS1 and DS2), Ixodes persulcatus(IP1 and IP2), Haemaphysalis concinna(HC1 and HC2) and Haemaphysalis japonica(HJ), respectively. The similarity of COⅠ sequences between IP1 and IP2 samples and Ixodes persulcatus published in GenBank No. JQ737101.1 and KU935457.1 was 99.85%-100.00%;the similarity of COⅠ sequences between HC1 and HC2 samples and Haemaphysalis concinna published in GenBank No. KR108861.1 was 99.84%;the similarity of COⅠ sequences between HJ sample and Haemophilus japonicus published in GenBank No. KR108879.1 was 99.84%, the above results were consistent with morphological identification. The similarity of COⅠ sequences between DS1 and DS2 samples and Dermacentor nuttalli published in GenBank No. KU594270.1 was 99.84%, the similarity of COⅠ sequences between DS1 and DS2 samples and Dermacentor silvarum published in GenBank No. MH079424.1 was 99.22%, although both of them met the criteria for direct identification, the samples were all confirmed to be Dermacentor silvarum based on the morphological characteristics of Dermacentor nuttalli and Dermacentor silvarum. Conclusion Based on the traditional morphological identification and classification, combined with DNA barcoding technology, the species of ticks can be identified accurately and rapidly.

关 键 词：黑瞎子岛 全沟硬蜱 森林革蜱 嗜群血蜱 日本血蜱 形态学种类鉴定 DNA条码技术 

分 类 号：R184.3[医药卫生—流行病学]