Application of DNA barcoding technique in identification of common flea species in plague foci of Hebei province, China
LAN Xiao-yu1, LU Liang2, HOU Zhi-lin1, DU Guo-yi1, SHI Xian-ming1, CUI Yao-ren1, LIU Guan-chun1, CHEN Yong-ming1, KANG Dong-mei1, ZHENG Nan1, REN Xing-yu1, YAN Dong1
1 Anti-plague Institute of Hebei Province, Zhangjiakou 075000, Hebei Province, China; 2 State Key Laboratory of Infectious Diseases Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention
Abstract：Objective To perform molecular biological identification of the common plague vector fleas in Hebei province, China by analyzing the mitochondrial cytochrome c oxidase subunitⅠ(COⅠ) gene and cytochrome c oxidase subunit Ⅱ:(COⅡ) gene, and to establish a DNA barcode database of common flea species in Hebei province. Methods Rodent-parasitic fleas and burrow fleas were collected in Kangbao ranch of Zhangjiakou, Hebei province from June to August, 2018, and rodent-parasitic fleas were collected in Chongli district of Zhangjiakou from September to October, 2018. After morphological identification, total DNA was extracted from various species of fleas of intact specimens, and then the mitochondrial COⅠ and COⅡ gene fragments were amplified and sequenced. The sequences were compared by BLAST for homology with those of fleas in GenBank. The phylogenetic tree of the COⅠ gene and COⅡ gene sequences was constructed by the neighbor-joining method. Results The specific COⅠ and COⅡ gene bands were amplified by PCR in all 16 samples of fleas. Among them, the COⅠ gene was sequenced successfully in 13 samples, and the COⅡ gene in 12 samples. According to the results of molecular phylogenetic tree of the COⅠ gene, 12 samples were consistent with morphological identification, and 1 sample was not. The results of molecular phylogenetic tree of the COⅡ gene were all consistent with the morphological identification in 12 samples. Conclusion Both COⅠ and COⅡ genes can be used as DNA barcode markers for species identification of common fleas in plague foci of Hebei province. Simultaneous identification of fleas using two genes improves the success rate of identification. The accumulated data will help establish an available database for future flea molecular identification technology.
兰晓宇, 鲁亮, 候芝林, 杜国义, 史献明, 崔耀仁, 刘冠纯, 陈永明, 康东梅, 郑楠, 任兴宇, 闫东. DNA条形码技术在河北省鼠疫疫源地常见蚤种鉴定中的应用[J]. 中国媒介生物学及控制杂志, 2020, 31(6): 662-666.
LAN Xiao-yu, LU Liang, HOU Zhi-lin, DU Guo-yi, SHI Xian-ming, CUI Yao-ren, LIU Guan-chun, CHEN Yong-ming, KANG Dong-mei, ZHENG Nan, REN Xing-yu, YAN Dong. Application of DNA barcoding technique in identification of common flea species in plague foci of Hebei province, China. Chines Journal of Vector Biology and Control, 2020, 31(6): 662-666.
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2020, Vol. 31 
