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

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