Taq Man技术快速定量检测人巨细胞病毒的早期感染  

Rapid Quantitative Detection of Human Cytomegalovirus by TaqMan PCR Assay

作　　者：邹俊煊[1] 张楚瑜[1] 游上游[2] 金建年[2] 

机构地区：[1]武汉大学生命科学学院,湖北武汉430072 [2]武汉市儿童医院,湖北武汉430022

出　　处：《武汉大学学报（理学版）》2004年第2期247-250,共4页Journal of Wuhan University:Natural Science Edition

基　　金：国家自然科学基金资助(30371501)

摘　　要：采用实时TaqMan技术快速定量检测血清中的人巨细胞病毒(HCMV)感染,在HCMV保守性的Ma jorIE基因区段设计了一对引物和一条Taqman探针,在LightCycler及Rotor Gene定量检测仪上检测HCMVDNA.试验证明,两种仪器定量结果具有一致性,以PCR模板浓度来定义,敏感度达到1.0×102拷贝/μL,线性范围为1.0×102～1.0×108拷贝/μL.应用该方法与ELISA法对40例临床婴幼儿患者血清同时进行HCMV检测,结果显示TaqMan法与ELISA法相关性不高,前者更适于HCMV感染早期的定量检测,提示该法可应用于孕妇及新生儿HCMV筛查,并有助于疗效监测和评估.A real-time TaqMan assay was applied to quickly quantify human cytomegalovirus (HCMV) DNA from human serum. A pair of primes and a TaqMan probe were designed that are specific for the recognition of a conservative Major IE gene in HCMV genome. LightCycler and Rotor-Gene instruments were separately used to quantify HCMV DNA. According to our results of the two instruments, HCMV TaqMan assay has a sensitivity of 1.0×10~2 DNA copy/μL with a dynamic range of detection between 1.0×10~2 and 1.0×10~8 DNA copy/μL. In order to evaluate its performance in clinical diagnosis, TaqMan was used to examine the HCMV DNA levels in sera from 40 sero-positive or negative subjects. The quantification results indicate that the TaqMan assay has low correlation to ELISA and suggest that this approach can apply broadly to early diagnose HCMV-related infections as well as to monitor and evaluate the relevant treatment, especially for pregnant women and newborn children.

关 键 词：TAQMAN技术 定量检测 巨细胞病毒 早期感染 早期诊断 HCMV检测 

分 类 号：R446.1[医药卫生—诊断学]