可视化核酸试纸条法快速检测松材线虫

摘要

A rapid and visual method was applied to detect Bursaphelenchus xylophilus.A pair of special primers and one competitive oligo nucleotide primer were designed.The 5'end of the forward primer was labeled with biotin and reverse primer with FITC,respectively.The proper concentration of the competitive primer was adjusted to avoid false positive results of nucleic acid strip assay.Then this visual detection method of B.xylophilus was established successfully.This visual detection method was simple,specific,sensitive,visual,stable and reliable.It can be widely applied in rapid identification of B.xylophilus in entry-exit inspection and quarantine.%建立了快速、可视化检测松材线虫的方法,分别设计一对特异性引物和一条竞争引物,其中上、下游引物的5'端分别修饰有生物素(biotin)和荧光素(FITC)；通过试验筛选并确定了竞争引物的最适比例从而避免核酸试纸条检测扩增产物的假阳性结果,实现了松材线虫的可视化检测.该技术具有操作便利、特异性强、灵敏度高,结果直观、稳定、可靠等优点,在口岸检验检疫实验室快速鉴定松材线虫方面具有良好的应用前景.