冻存血浆中microRNAs稳定性研究  

Stability of microRNAs in frozen plasma

作　　者：韩璐璐[1] 高小晶[1] 管晓媛[1] 段兵[1] 黄一玲[1] 谢爽[1] 许莉[1] 刘红[1] 李一石[1] 

机构地区：[1]中国医学科学院 北京协和医学院 阜外心血管病医院 卫生部心血管药物临床研究重点实验室,北京市100037

出　　处：《中华实用诊断与治疗杂志》2012年第7期639-641,共3页Journal of Chinese Practical Diagnosis and Therapy

基　　金："重大新药创制"科技重大专项(2008ZX09312-018;2012ZX09303008-001)

摘　　要：目的探讨不同冻存时间血浆microRNAs表达水平及血浆总RNA提取并冻存2周后microRNAs表达水平的稳定性。方法 -80℃冻存2a,1a及6个月和新鲜抽取的健康人血浆样本各10份,提取血浆总RNA,应用基于TaqMan探针的MicroRNA定量检测法检测血浆样本中4种microRNAs表达水平。结果冻存6个月人血浆样本中U6与新鲜血浆标本比较差异有统计学意义(P<0.05),冻存2a,1a及6个月人血浆样本中4种microRNAs表达水平与新鲜血浆标本比较差异无统计学意义(P>0.05);新鲜血浆RNA提取并冻存2周后2个目的microRNAs相对表达水平与提取后即刻比较差异无统计学意义(P>0.05)。结论 -80℃下冻存2a内血浆中microRNAs稳定存在,血浆总RNA提取并冻存2周后仍可用于microRNAs的定量检测。Objective To evaluate the microRNAs expression levels in frozen plasma at different frozen time and the stability of mieroRNAs in total RNA extracted and frozen for 2 weeks. Methods MirVana PARIS Kit was used to isolate total RNA from plasma frozen at-80 ℃ for 2 years, 1 years and 6 months and plasma of 10 healthy individuals. The expression levels of 4 mieroRNAs were detected with TaqMan microRNA assays quantitative real-time PCR. Results There were no significant differences in the levels of the same mieroRNAs between fresh plasma and plasma frozen for 2 years, 1 year or 6 months except U6 in plasma frozen for 6 months （P〉0.05）. There was no significant difference between microRNAs relative expression levels being isolated for 2 weeks RNA and being isolated instantly （P〉0.05）. Conclusion MicroRNAs in plasma frozen at -80 ℃ for 2 years are stable. Total RNA extracted and isolated for 2 weeks can be used for quantitative detection of microRNAs.

关 键 词：血浆 MICRORNA 冻存 QRT-PCR 

分 类 号：R966[医药卫生—药理学]