CAJ | 학술논문

目的探讨多重荧光定量PCR技术的优化条件,建立基于AllGlo探针技术荧光定量法检测4种疱疹病毒新方法.方法分别采用单重和多重定性PCR扩增临床常见4种疱疹病毒[单纯疱疹病毒1型(HSV-1)、HSV-2、Epstein-Barr病毒(EBV)、巨细胞病毒(CMV)]并测序鉴定,然后分别采用AllGlo探针和TaqMan探针的单重和多重定量PCR技术对4种疱疹病毒进行单种和多种病毒同时定性定量检测.结果 TaqMan探针和AllGlo探针单种疱疹病毒检测阳性率和特异性均为100％,AllGlo探针单重定量PCR检测比4重探针单种定量PCR的检测Ct值高1～3,AllGlo探针4重定量PCR可以同时检测4种疱疹病毒,相同样品AllGlo探针4重定量PCR检测与单探针单重定量PCR分别检测的结果符合率100％.结论 AllGlo探针荧光定量PCR技术的通量、灵敏度和特异性均高于TaqMan探针,应用前景广阔.
목적 탐토다중형광정량PCR기술적우화조건,건립기우AllGlo탐침기술형광정량법검측4충포진병독신방법.방법 분별채용단중화다중정성PCR확증림상상견4충포진병독[단순포진병독1형(HSV-1)、HSV-2、Epstein-Barr병독(EBV)、거세포병독(CMV)]병측서감정,연후분별채용AllGlo탐침화TaqMan탐침적단중화다중정량PCR기술대4충포진병독진행단충화다충병독동시정성정량검측.결과 TaqMan탐침화AllGlo탐침단충포진병독검측양성솔화특이성균위100％,AllGlo탐침단중정량PCR검측비4중탐침단충정량PCR적검측Ct치고1～3,AllGlo탐침4중정량PCR가이동시검측4충포진병독,상동양품AllGlo탐침4중정량PCR검측여단탐침단중정량PCR분별검측적결과부합솔100％.결론 AllGlo탐침형광정량PCR기술적통량、령민도화특이성균고우TaqMan탐침,응용전경엄활.
Objective To optimize the condition of multiple fluorescence quantitative PCR and establish a new assay of four human herpes virus (HHV) detected by AllGlo quadruple fluorescence quantitative PCR.Methods Four HHV including HSV-1,HSV-2,EBV and CMV were identified by sequence analysing the qualitative PCR production.Furthermore,they were quantitatively detected by AllGlo and TaqMan multiple fluorescence quantitative PCR respectively.Results Both the positive rate and specificity of AllGlo and TaqMan in detecting single HHV achieved 100％.And AllGlo single fluorescence quantitative PCR prevailed over TaqMan's by Ct of 1-3.Four HHV can be simultaneously detected by AllGlo quadruple fluorescence quantitative PCR,comparing to the only two by TaqMan.Conclusion AllGlo fluorescence quantitative PCR assay allows a higher throughput,sensitivity and specificity than TaqMan in detection and thus provides a board prospect.