Chines Journal of Vector Biology and Control ›› 2014, Vol. 25 ›› Issue (1): 21-23.DOI: 10.11853/j.issn.1003.4692.2014.01.006

• Original Reports • Previous Articles     Next Articles

Application of DNA barcoding technique in identification of rodentspecies in natural focus of plague in Hebei province, China

YAN Dong1, LU Liang2, JIN Sheng-hao1, SHI Xian-ming1, CUI Yao-ren1, LIU Guan-chun1, LI Yu-wei2,LIU Qi-yong2, ZHENG Nan1, KANG Dong-mei1, BAI Xue-wei1, NIU Yan-fen1,CHEN Yong-ming1, ZHOU Song1, LAN Xiao-yu1, LI Zhen-hai1   

  1. 1 Institute of Plague Prevention and Control of Hebei Province, Zhangjiakou 075000, Hebei Province, China;
    2 State Key Laboratory for Infectious Diseases Prevention and Control, National Institute for CommunicableDisease Control and Prevention, Chinese Center for Disease Control and Prevention
  • Online:2014-02-20 Published:2014-02-20
  • Supported by:
    Supported by the Special Research Program for Health (No. 201202021) and Hebei Provincial Medical Science Research Foundation (No. 20110267)

DNA条形码技术在河北省鼠疫疫源地鼠种鉴定中的应用 

闫东1,鲁亮2,金圣浩1,史献明1,崔耀仁1,刘冠纯1,李玉伟2,刘起勇2,郑楠1,康东梅1,白雪薇1,牛艳芬1,陈永明1,周松1,兰晓宇1,李振海1   

  1. 1 河北省鼠疫防治所流行病科,河北 张家口 075000;
    2 中国疾病预防控制中心传染病预防控制所,传染病预防控制国家重点实验室 
  • 作者简介:闫东, 男, 主管医师, 从事鼠疫流行病研究工作。Email: yandong0000@126.com
  • 基金资助:
    卫生行业科研专项(201202021); 河北省医学科学研究重点课题(20110267)

Abstract: Objective To evaluate the feasibility of DNA barcoding in identifying the rodent species based on the common rodents in the natural focus of plague in Hebei province, China. Methods The rodent liver specimens were collected in the natural focus of plague in Hebei in 2012; the whole rodents were conserved, and then the stuffed specimens were made; genomic DNA was extracted from the rodent liver. The mitochondrial COⅠ gene was amplified by consensus primer PCR and then sequenced. BLAST comparison was made between the sequencing results and the DNA bar codes of other rodent species in GenBank, and molecular evolutionary tree was constructed. Results In this research, specific COⅠ gene bands were obtained by PCR amplification in all 87 samples. Among all samples, 85 had the molecular evolutionary tree in accordance with the results of morphological identification, and 2 had different results, but the field identification was found to be incorrect by identifying the skulls and morphological specimens repeatedly. Conclusion DNA barcoding technique can be used to effectively identify the rodent species. At the same time, it can also be used for discussing the phylogenetic relationship of rodent genus and species classification.

Key words: Plague natural focus, DNA barcoding, COⅠ gene

摘要: 目的 以河北省鼠疫疫源地常见鼠为对象,探讨DNA条形码技术在鼠类鉴定方面的可行性。方法 2012年在河北省鼠疫疫源地内采集鼠肝脏标本,并保存整只鼠,制作剥制标本,提取基因组DNA,用通用引物PCR 法扩增线粒体COⅠ基因,并测序。将测序结果与GenBank中其他鼠类物种的DNA条形码进行BLAST 比对,并构建分子进化树。结果87份样本均能通过PCR 扩增出特异性COⅠ基因条带,其中85份样本所构建的分子进化树结果与形态学鉴定结果一致,2份样本结果有差异,经反复鉴定头骨及形态标本,发现为现场鉴定错误。结论 DNA条形码技术能够对鼠类进行有效的物种鉴定,同时还可以探讨啮齿动物属、种分类单元的系统发育问题。

关键词: 鼠疫疫源地, DNA条形码, COⅠ基因

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