犬巴贝西虫病PCR检测方法的建立

摘要

One pair of specific primers was designed according to the 18sRNA gene sequence of canine Babesia,and then a PCR assay was established for detecting canine Babesia. A fragment in length of 319 bp was amplified by PCR and the sequence homology to those from GenBank was 100%. No PCR products were amplified from .purified DNA of Eperythrozoon or Pasteurella. This method can detect the genomic DNA of Canine Babesia in 10-3 diluted whole blood DNA of infected dogs. This indicates that the PCR method was precise,sensitive and specific.%为快速、准确检测犬巴贝西虫病,根据GenBank中收录的犬的巴贝西虫18sRNA基因序列,设计合成1对特异性诊断引物,建立了PCR诊断方法.结果扩增出了犬巴贝西虫18sRNA基因的一段大小为319bp片段,经测序,所扩增序列与报道的序列完全匹配.同时对附红细胞体、巴氏杆菌DNA样品进行扩增没有出现扩增条带.阳性犬全血DNA稀释1000倍后,依然可以有效地检测出虫体DNA.结果表明,所建立的PCR检测方法,具有极高的敏感性和特异性,可以运用于临床检测巴贝西虫病.