DNA条形码技术在兰州中川机场鼠类鉴定中的应用  

Application of DNA barcoding in classification of rodents at Lanzhou Zhongchuan airport area

作　　者：陈利伟 赵志亮 邱德义 巩伟兵 王莉 高晓强 刘芳 岳巧云 CHEN Li-wei, ZHAO Zhi-liang, QIU De-yi, GONG Wei-bing, WANG Li, GAO Xiao--qiang, LIU Fang, YUE Qiao-yun(Zhongshan Entry-Exit Inspection and Quarantine Bureau Technology Center, Zhongshan, Guangdong 528403, Chin)

机构地区：[1]中山出入境检验检疫局检验检疫技术中心,广东中山528403 [2]广东药科大学公共卫生学院 [3]甘肃国际旅行卫生保健中心 [4]中山大学生命科学学院

出　　处：《中国国境卫生检疫杂志》2018年第2期102-105,共4页Chinese Journal of Frontier Health and Quarantine

基　　金：国家重点研发计划项目(2016YFF0203205);甘肃省科技支撑计划项目(1604FKCA121);甘肃出入境检验检疫局科技计划项目(2014GSYJ01)

摘　　要：目的调查兰州中川机场及其附近地区鼠类的组成,并利用DNA条形码技术对鼠种进行鉴定。方法 2015年1月—2017年7月,在中川机场及其附近地区采用笼夜法采集鼠类,提取鼠肾DNA,用PCR方法扩增线粒体细胞色素c氧化酶亚基Ⅰ(COⅠ)基因并测序,测序结果在DNA条形码专用数据库BOLD中进行比对,确定鼠种。利用Mega 6.0构建分子进化树。对于形态学和COⅠ基因鉴定结果不一致的样本,进一步通过细胞色素b(Cytb)基因进行分子鉴定。结果 118份样品均能扩增出特异性COⅠ基因片段,115份样本的分子鉴定结果与形态学结果基本一致,3份样本形态学鉴定结果与分子鉴定有差异,经Cytb基因序列比对及形态学反复鉴定,确定初步形态学鉴定结果有误,采用分子鉴定结果。结论 DNA条形码技术可以对鼠类进行有效鉴定,是对形态学鉴定的有力补充,为口岸病媒生物的快速识别和鉴定提供了科学依据。Objective To investigate the composition of rodents in Lanzhou Zhongchuan airport areas,and identify the rodents with DNA barcoding. Methods The rodents were collected by cage-trap overnight at Lanzhou Zhongchuan airport areas during January 2015 to July 2017. The genomic DNA was extracted from the rodent kid-ney samples. The mitochondrial cytochrome c oxidase subunit Ⅰ gene（CO Ⅰ） was amplified by PCR and then sequenced. The sequences were aligned in the dedicated DNA barcode database-BOLD system,and a phylogenetic tree was constructed by Mega 6.0 software. For the samples whose morphological identification were inconsistent with the molecular identification based on CO Ⅰ gene,Cytb gene was further used to analyzed. Results The specific CO Ⅰsequences were successfully amplified from 118 rodents by PCR. Molecular identification were identical with those of morphological identification for the 115 out of the 118 samples,for those 3 with different results,the initial morphological identification was found to be incorrect after Cytb gene sequence alignment and morphological identification,molecular identification was adopted finally. Conclusion DNA barcoding can be used as an effective technique to identify rodent species,it is a powerful complement to the morphological identification,and it provides a scientific basis for the rapid identification of medical vectors.

关 键 词：DNA条形码 细胞色素C氧化酶亚基Ⅰ基因 鼠类 

分 类 号：R184.35[医药卫生—流行病学] S443[医药卫生—公共卫生与预防医学]