实时RT-PCR检测二氢嘧啶脱氢酶mRNA的表达  

Determination of dihydropyrimidine dehydrogenase expression by real-time RT-PCR

作　　者：陆志成[1] 蔡刚[2] 沈茜[2] 

机构地区：[1]上海市第七人民医院,上海200137 [2]第二军医大学附属长海医院检验科,上海200433

出　　处：《检验医学》2005年第1期59-62,共4页Laboratory Medicine

摘　　要：　　目的　建立实时逆转录聚合酶链反应(RT PCR)检测二氢嘧啶脱氢酶mRNA的定量方法,并分别从 RNA水平与蛋白质水平检测二氢嘧啶脱氢酶在人体部分组织中的表达。方法　采用基于TaqMan探针技术的 实时RT PCR方法检测二氢嘧啶脱氢酶mRNA的表达,用酶联免疫吸附试验(ELISA)方法检测其蛋白质表达水 平。结果　实时RT PCR检测二氢嘧啶脱氢酶的线性范围可达102～109拷贝数/μl,批内误差<5%。结论　这 一实时RT PCR的方法可以从纳克级的总RNA中检出二氢嘧啶脱氢酶的表达,并且ELISA结果与实时RT PCR 结果有较好的相关性。Objective To establish the real-time RT-PCR method to detect the mRNA level of dihydropyrimidine dehydrogenase, and to determine the expression of dihydropyrimidine dehydrogenase in various human tissues in RNA level and protein level. Methods The mRNA expression level of dihydropyrimidine dehydrogenase was detected by TagMan real-time RT-PCR. Dihydropyrimidine dehydrogenase protein level was measured by ELISA. Results The detection range of real-time RT-PCR was 102-109 copy number/μl. The standard error within sample was less than 5%. Conclusions The real-time RT-PCR method can detect the mRNA of dihydropyrimidine dehydrogenase in the level of several ng of total RNA. The results of ELISA are correlated with that of the real-time RT-PCR.

关 键 词：二氢嘧啶脱氢酶 蛋白质表达 总RNA RT-PCR检测 酶联免疫吸附试验(ELISA) 实时逆转录聚合酶链反应 体部 拷贝数 TAQMAN探针 蛋白质水平 

分 类 号：R730[医药卫生—肿瘤] R979.1[医药卫生—临床医学]