猫细小病毒 PCR 检测方法的建立及初步应用

摘要: 目的 建立猫细小病毒 PCR 检测方法，应用于猫临床样本中 FPV 的快速检测。方法 根据已发表的 FPV VP2 基因序列设计合成引物，并以此建立 FPV 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 33 份猫临床样品进行检测。结果 建立的 FPV PCR 检测方法与猫疱疹病毒Ⅰ型(FHV-1)、猫冠 状病毒(FeCV)、猫合胞体病毒(FeSFV)、猫免疫缺陷病毒(FIV)均无交叉反应;可检测病毒最小滴度为 5lgTCID50 / mL，相应的 DNA 模板浓度为 4. 9 × 102 拷贝/μL;FPV DNA 在 － 30℃冰箱放置 12 个月仍可检测出目的条带。应用 该方法从 33 份猫临床样本中检测出 21 份 FPV 核酸阳性。结论 建立的 FPV PCR 检测方法具有特异、敏感及稳定 的特点，适合于临床 FPV 的感染检测。

Establishment and Preliminary Application of PCR Method for Detection of Feline Panleukopenia Virus

Abstract:Objective To establish a PCR method for detection of Feline panleukopenia virus ( FPV) in cats． Method The primers were designed and synthesized according to the published FPV specific sequences of VP2 gene． PCR method was established，and the specificity，sensitivity and stability of this method were tested． Then the method was used to detect 33 clinical samples of cats． Result The developed PCR method was no cross reaction with Feline herpes virus 1 ( FHV-1 )，Feline coronavirus ( FeCV)，Feline syncytium-forming virus (FeSFV)，Feline immunodefiency virus(FIV)． Its minimum detection limit (the lowest detection virus titer) was 5lgTCID50 /mL，the corresponding template DNA concentration was 4. 9 x 102 copies． FPV DNA，maintained at － 30 ℃ for 12 months，was still able to amplify the fragment of about 301 bp． The 33 cats were 21 positive ，and 11 negative for FPV detected by PCR． Conclusion The developed PCR method is good in specificity，sensitivity， stability，and can be used for detecting the FPV in cat．