实验动物科学 ›› 2022, Vol. 39 ›› Issue (5): 69-71.DOI: 10. 3969 / j. issn. 1006-6179. 2022. 05. 013

• 技术 • 上一篇    下一篇

环介导等温扩增技术检测四翼无刺线虫方法的建立

  

  1. (中国食品药品检定研究院实验动物资源研究所,北京 102629)
  • 收稿日期:2019-09-30 出版日期:2022-10-28 发布日期:2022-11-15
  • 通讯作者: 岳秉飞( 1960—) ,男,研究员,博士. 研究方向:动物遗传学. E-mail:y6784@ 126. com
  • 作者简介:黄 健( 1988—) ,女,助理研究员,研究方向:实验动物寄生虫学检测. E-mail:huangjian88hj@ 126. com
  • 基金资助:
    国家重点研发计划资助( 2017YFD0501600) ;国家重点研发项目省级资金资助(GX18B052) 。

Establishment of Loop-mediated Isothermal Amplification Method for Aspiculuris Tetraptera

  1. ( Institute for Laboratory Animal Resources,National Institute for Food and Drug Control, Beijing 102629, China)
  • Received:2019-09-30 Online:2022-10-28 Published:2022-11-15

摘要: 目的 建立快速高效的鉴定四翼无刺线虫的环介导等温扩增方法( LAMP) 。 方法 根据四翼无刺线虫已发表的 COI 序列设计 LMAP 引物,以鞭毛虫和水为对照,进行特异性评价。 四翼无刺线虫基因组 DNA 梯度稀释后,验证其敏感性。 通过荧光显色和琼脂糖凝胶电泳对 LAMP 产物进行鉴定。 结果 LAMP 产物经荧光显色后观察结果,阳性为绿色,阴性为棕色。 四翼无刺线虫的 LAMP 产物经电泳后呈现特征性的梯状条带,对照组均无条带。结论 成功建立四翼无刺线虫 LAMP 检测方法。

关键词: 四翼无刺线虫, 环介导等温扩增技术, 检测

Abstract: Objective To establish a rapid and efficient loop mediated isothermal amplification method for aspiculuris tetraptera. Method The LMAP primers were designed according to the published COI sequence of aspiculuris tetraptera, and Giardia lamblia and water were used as controls for specificity evaluation. The genomic DNA of aspiculuris tetraptera was diluted in series to verify its sensitivity. LAMP products were identified by fluorescent color and agarose gel electrophoresis. Result The LAMP product is colored by fluorescence and the result is green, and the brown is negative. After electrophoresis, the LAMP products of aspiculuris tetraptera showed characteristic ladder-like bands. There was no band in the control group. Conclusion A simple method for detecting aspiculuris tetraptera by LAMP assay was established.

Key words: aspiculuris tetraptera, loop mediated isothermal amplification, detection

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