根据GenBank上公布的简单异尖线虫核糖体DNA的内转录区(ITS-1、5.8S、ITS-2)序列设计特异性引物,对线虫样品进行PCR扩增并将产物克隆到pMD18-T载体后测序。结果表明,扩增的目的片段大小为430bp,与预期扩增序列同源性为99%。该PCR检测体系的特异性强,不能在宫脂线虫、伪地新线虫DNA中扩增出条带;敏感性高,最低检测的DNA含量为0.4pg。该检测体系的建立为简单异尖线虫的检测、鉴定和流行病学调查提供了有力的技术支持。 Specific primers were designed according to ITS-1, 5.8S, ITS-2 sequences published on GenBank. The nematode samples were PCR amplified and cloned into pMD18-T vector After sequencing. The results showed that the size of the amplified fragment was 430bp, which was 99% homologous to the expected amplified sequence. The PCR detection system is highly specific and can not amplify bands in the genome of C. occidentalis and P. pseudoaneuria. The sensitivity is high, and the minimum detectable DNA content is 0.4 pg. The establishment of the detection system provides a powerful technical support for the detection, identification and epidemiological investigation of Isoprenia simplex.