DNA条形码技术应用于口岸吸血蠓类快速分子鉴定的研究  

Study on DNA barcoding technology applied in rapid molecular identification of bloodsucking midges in port

作　　者：贺骥 宋福春[2] 苏水宽 王长军[3] 陈剑 陈慧 冯梅 傅健 

机构地区：[1]厦门国际旅行卫生保健中心,福建厦门361012 [2]安徽理工大学医学院,安徽淮南232001 [3]南京军区疾病预防控制中心,江苏南京210002

出　　处：《中华卫生杀虫药械》2014年第2期131-135,共5页Chinese Journal of Hygienic Insecticides and Equipments

基　　金：国家质检总局科研项目(编号:2012IK242;2012IK244);厦门市科研项目(编号:3502Z20124005)

摘　　要：目的建立口岸吸血蠓类快速分子鉴定方法，提高鉴定效率和准确率。方法选择口岸区域采集点，采集吸血蠓类。经形态学鉴定后，单只蠓提取基因组DNA，采用自行设计的COI基因扩增引物进行扩增。PCR产物经电泳鉴定后，进行双向测序。序列经分析后，去除引物序列和不稳定序列，进行拼接。提交至NC—BI和BOLD数据库进行比对，并建立系统进化树进行验证。结果得到12个新序列，已登录至GenBank，登录号为KF528689～KF528700。其中台湾蠛蠓的COI序列（683bp）填补了NCBI数据库的空白。所得序列经分析后可以将多种蠛蠓、库蠓很好地区分开。结论本项目所建立的基于吸血蠓类COI基因的DNA条形码快速分子鉴定方法可以很好地鉴定各吸血蠓种，值得进一步推广应用。Objective To improve the efficiency and accuracy of identification,establish the method for rapid molec- ular identification of bloodsucking midges in ports. Methods Selecting sampling points in ports, and collecting the bloodsucking midges. After morphological identification, the genomic DNA was extracted from single midge. The cyto- chrome oxidase subunit I （COI） gene was amplified with primers designed by us. After electrophoretic analysis, the PCR products were sequenced bidirectionally. After sequences analysis, there were the primer sequences and instabili- ty sequences. The sequences were submitted to the database of NCBI and BOLD. Results 12 new sequences were sent to the GenBank （ accession no. KF528689 - KF528700）. The COI sequence of Lasiohelea taiwana filled the gaps of the NCBI database. Sequences obtained could distinguish many bloodsucking midges. Conclusion The rapid molecular identification method using DNA barcoding based bloodsucking midges COI gene can be applied to identify various species of blood-sucking midges.

关 键 词：COI DNA条形码 吸血蠓 分子鉴定 口岸 

分 类 号：R384.5[医药卫生—医学寄生虫学]