常规TRIzol法提取血清microRNA的改良  

Optimization of microRNA extraction method from serum

作　　者：聂宇[1] 张洋[2] 韩变梅[1] 王恺隽[2] 丛祥凤[2] 陈曦[1] 

机构地区：[1]北京协和医学院中国医学科学院阜外心血管病医院卫生部心血管疾病再生医学重点实验室,北京市100037 [2]北京协和医学院中国医学科学院阜外心血管病医院临床检验中心,北京市100037

出　　处：《中华实用诊断与治疗杂志》2011年第5期427-428,432,共3页Journal of Chinese Practical Diagnosis and Therapy

基　　金：973重大基础专项(2010CB529500);国家自然科学基金(30770862);首都医学发展基金(2009-2013)

摘　　要：目的:对现有常规提取血清microRNA的方法进行改良。方法:运用常规TRIzol法和改良后的血清microRNA提取方法分别提取健康人血清中的microRNA;利用反转录茎环引物及TaqMan探针进行实时荧光定量PCR检测miR-16和miR-21的含量,通过比较miR-16和miR-21的Ct值以鉴定不同提取方法对microRNA检测的影响。结果:改良后的血清microRNA提取方法可明显降低PCR检测中miR-16和miR-21的Ct值。结论:改良的血清microRNA提取方法可有效提高目的microRNA的检出水平。Objective To optimize the method of extracting microRNA from serum.Methods MicroRNA was extracted from serum with the common and the optimized methods respectively.The contents of miR-16 and miR-21 were detected with TaqMan qRT-PCR technique.Ct values were compared to evaluate the influence of different methods on microRNA detection.Results Ct values of miR-16 and miR-21 in PCR amplification were significantly reduced when microRNAs were extracted with optimized method.Conclusion The optimized method of extracting microRNA from serum improves the sensitivity of detection of microRNA significantly.

关 键 词：血清microRNA RNA提取技术 TaqManqRT-PCR 

分 类 号：R730.2[医药卫生—肿瘤]