Taq Man技术快速定量检测人巨细胞病毒的早期感染

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 邹俊煊 ZOU Jun-xuan~1 1 2 3 4 first-author 张楚瑜 ZHANG Chu-yu~1 1 2 3 4 游上游 YOU Shang-you~2 1 2 3 4 金建年 JIN Jian-nian~2 1 2 3 4 武汉大学生命科学学院 武汉大学生命科学学院 College of Life Sciences, Wuhan University College of Life Sciences, Wuhan University 武汉市儿童医院 武汉市儿童医院 Wuhan Children’s Hospital Wuhan Children’s Hospital 武汉市儿童医院,湖北,武汉,430072 武汉市儿童医院,湖北,武汉,430072 ,湖北,武汉,430022 ,湖北,武汉,430022 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. 人巨细胞病毒 TaqMan 定量检测 早期诊断 human cytomegalovirus（HCMV） TaqMan quantitative detection early diagnosis R446.5 国家自然科学基金资助（30371501） 2004-02-01 2021-04-01 2