热带病与寄生虫学

热带病与寄生虫学 ›› 2021, Vol. 19 ›› Issue (2): 101-103,111.

• 实验研究 • 上一篇    下一篇

基氏蠊螨的 18S rDNA 分子鉴定

李梦珠,张兰香,詹雨娟,褚凌渺,胡婷婷,李心玫,王严,孙恩涛
  

  1. 皖南医学院检验学院,安徽 芜湖 241002
  • 出版日期:2021-04-20 发布日期:2021-04-22
  • 通信作者: 孙恩涛,E-mail:asdentao@ 126. com
  • 作者简介:李梦珠,女,本科在读,研究方向:媒介生物检验与检疫。 E-mail:2125797941@ qq. com
  • 基金资助:
    国家自然科学基金项目(31870352);皖南医学院 2019 年青年卓越人才资助项目(WYQNYX201902);安徽省省级大学生创新创业项
    目    (S202010368034、S202010368104)

Molecular identification of Blattisocius keegani based on 18S rDNA gene sequences 

LI Meng-zhu,ZHANG Lan-xiang,ZHAN Yu-juan,CHU Ling-miao,HU Ting-ting,LI Xin-mei,WANG Yan,SUN En-tao    

  1. School of Laboratory Medicine,Wannan Medical College,Wuhu 241002,Anhui Province,China
  • Online:2021-04-20 Published:2021-04-22

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摘要： 目的 基于核糖体 18S rDNA 基因对基氏蠊螨进行分子鉴定。 方法 采集和分离储藏物样本,进行螨类的形态学鉴定。 提取单个螨的基因组 DNA,经 PCR 扩增、克隆和测序获得 COⅠ基因和 18S rDNA 基因,将所获序列进行 Blast 对比。 检索 GenBank 数据库中蠊螨属 18S rDNA 基因序列,利用 Clustal X 1. 83 软件进行多序列比对,基于MEGA X 软件进行序列分析并以邻接法(Neighbor-Joining,NJ)构建系统发育树。 结果 采集的样本经形态和 COⅠ基因双重鉴定为基氏蠊螨。 同时,所选取的 10 个基氏蠊螨的 18S rDNA 基因序列完全一致,均表现出 A / T 碱基偏向性,与同属的 Blattisocius tarsalis 和 Blattisocius everti 分别有 98. 73%和 98. 94%的同源性。 基于 18S rDNA 基因序列的系统进化树显示,基氏蠊螨与 Blattisocius tarsalis 和 Blattisocius everti 聚为一支。 结论 本研究建立了基氏蠊螨基于 18S rDNA 基因序列的分子鉴定方法,为基氏蠊螨的准确鉴定奠定基础。

关键词: 基氏蠊螨, 分子鉴定, 核糖体 18S rDNA

Abstract: Objective To establish a molecular method for identification of Blattisocius keegani based on ribosomal 18S rDNA gene sequences. Methods Storage samples were collected and isolated. Genomic DNA of a single mite was extracted on morphological identification basis. The COⅠand 18S rDNA gene sequences were obtained by PCR amplification,cloning and sequencing. Then the obtained sequences were aligned using Blast software,and 18S rDNA sequences of Blattisocius were retrieved from the GenBank. Multiple sequence alignment was done using the software Clustal(version 1. 83) with the known gene sequences. MEGA X software was used to analyze sequences,and the phylogenetic tree were constructed by neighbor-joining(NJ) method. Results The mites were identified as Blattisocius keegani based on COⅠ and morphological identification. The amplified sequences of the ten Blattisocius keegani were identical,showing A / T bias. The sequence homology of Blattisocius keegani with Blattisocius tarsalis and Blattisocius everti was 98. 73% and 98. 94%,respectively. Phylogenetic analysis based on 18S rDNA genes showed that the three mites were clustered into one branch. Conclusion We have successfully established a method for identifying Blattisocius keegani  on 18S rDNA sequences basis,which can be a foundation for accurate identification of Blattisocius keegani .

Key words: Blattisocius keegani , Molecular identification, Ribosomal 18S rDNA

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