CAJ | 학술논문

目的：建立猫疱疹病毒I型（ FHV-1）实时荧光定量PCR检测方法，应用于实验用猫及临床患猫FHV-1的快速检测。方法根据已发表的FHV-1 TK基因序列设计特异引物和TaqMan探针，使用分子生物学方法制备质粒标准品，建立FHV-1实时荧光定量PCR方法，并对方法的线性、特异性、敏感性、及稳定性等进行测定。用建立的方法对48份猫临床样品进行检测。结果成功建立FHV-1实时荧光定量PCR方法；该方法的线性范围为（1×102～1×109）copies／μL，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，检测灵敏度可达到10 copies／μL。批内变异系数均小于5％。应用该方法从48份猫临床样本中检测出33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的快速定量检测。
목적：건립묘포진병독I형（ FHV-1）실시형광정량PCR검측방법，응용우실험용묘급림상환묘FHV-1적쾌속검측。방법근거이발표적FHV-1 TK기인서렬설계특이인물화TaqMan탐침，사용분자생물학방법제비질립표준품，건립FHV-1실시형광정량PCR방법，병대방법적선성、특이성、민감성、급은정성등진행측정。용건립적방법대48빈묘림상양품진행검측。결과성공건립FHV-1실시형광정량PCR방법；해방법적선성범위위（1×102～1×109）copies／μL，여단순포진병독Ⅰ형（HSV-1）、견포진병독（CHV）、저위광견병독（PRV）、묘세소병독（ FPV）균무교차반응，검측령민도가체도10 copies／μL。비내변이계수균소우5％。응용해방법종48빈묘림상양본중검측출33빈FHV-1핵산양성。결론건립적FHV-1PCR검측방법구유특이、민감급은정적특점，괄합우림상FHV-1적쾌속정량검측。
Objective To establish a real-time fluorescent quantitative PCR ( Q-PCR) method for detection of feline herpesvirus 1 ( FHV-1 ) in experiment cats and clinical sick cats.Methods Primers and TaqMan probes were designed and synthesized according to the published FHV-1 specific sequences of TK gene.FHV DNA standards were prepared using molecular biological techniques.The linearity, specificity, sensitivity, stability of the established Q-PCR method were tested.The method was used to detect 48 samples of cats.Results The linear range was 102 copies/μL to 109 copies/μL.The developed Q-PCR method showed no cross reaction with herpes virus type 1 ( HSV-1 ) , canine herpesvirus (CHV), pig pseudo rabies virus (PRV) and cat parvovirus (FPV).The sensitivity was 10 copies/μL.The coefficient of variation ( CV ) was less than 5%.There were 33 positive cases detected in the 48 samples of cats. Conclusions The developed Q-PCR method is good in linearity, specificity, sensitivity, stability, and may be used for rapid quantitative detection of FHV-1 in cats.