猫疱疹病毒Ⅰ型PCR检测方法的建立及初步应用

目的建立猫疱疹病毒Ⅰ型（FHV-1）PCR检测方法,应用于临床样品中FHV-1的快速检测。方法根据已发表的FHV-1TK基因序列设计合成引物,并以此建立FHV-1的PCR检测方法,并对方法的特异性、敏感性、稳定性等进行验证。用建立的方法对15份猫临床样品进行检测。结果建立的FHV-1PCR检测方法与单纯疱疹病毒Ⅰ型（HSV-1）、犬疱疹病毒（CHV）、猪伪狂犬病毒（PRV）、猫细小病毒（FPV）均无交叉反应;可检测病毒最小滴度为7lg TCID50/ml,相应的DNA模板浓度为1.46×102拷贝/ml;FHV-1DNA在-30℃冰箱放置12个月仍可检测出目的条带。应用该方法从15份猫临床样本中检测出10份FHV-1核酸阳性。结论建立的FHV-1PCR检测方法具有特异、敏感及稳定的特点,适合于临床FHV-1的感染检测。

Establishment and preliminary application of PCR method for detection of felid herpesvirus

Objective To establish a PCR method for the detection of felid herpesvirus 1（FHV-1）in cat.Methods Primers were designed and synthesized according to the published FHV-1specific sequences of TK gene.PCR method was established,and the specificity,sensitivity,repeatability of this method were evaluated.The established method was used to detect samples from 15 cats.Results The developed PCR method was tested with no cross reactions with herpes simplex virus 1（HSV-1）,Canine herpesvirus（CHV）,Porcine pseudorabies virus（PRV）,and Feline panleukopenia virus（FPV）.The lowest detection virus titer is 7lg TCID50/ml,the corresponding template DNA concentration was 1.46×102 copies;FHV-1DNA maintained at-30℃for 12 months can still be detected.10 out of 15 cats were positive by PCR.Conclusions An efficient PCR method was established to detect FHV-1in cat.