实时RT-PCR常用内对照在死亡早期人心肌内的稳定性  

Evaluation on Stability of Internal Controls in Human Cardiac Muscle by Real-Time RT-PCR during Early Postmortem Interval

作　　者：张萍[1] 马开军 张恒[1] 王慧君[3] 沈忆文[1] 陈龙[1] 

机构地区：[1]复旦大学上海医学院法医学系,上海200032 [2]上海市公安局物证鉴定中心 上海市现场物证重点实验室,上海200043 [3]复旦大学附属儿科医院儿童发育与疾病转化医学研究中心,上海201102

出　　处：《法医学杂志》2012年第2期81-84,共4页Journal of Forensic Medicine

基　　金：国家自然科学基金资助项目(81172896)

摘　　要：目的探讨实时RT-PCR方法常用内对照表达水平在死亡早期人体心肌组织中的稳定性。方法选取10例具有一致的外界环境(平均存放温度25℃)、不同死亡时间(4.3～22.3 h)的个体,提取心肌组织的总RNA。选择6个常用内对照:β-actin、GAPDH、B2M、U6、18S rRNA、HSA-miR-1,通过实时RT-PCR检测其在心肌组织的表达水平。利用genormPLUS软件评估内对照在死亡早期表达水平的稳定性,挑选出最稳定的内对照并且分析其表达水平与相关因素(年龄、性别、死因)的关系。结果死亡早期心肌组织中U6表达水平最稳定,其表达量与年龄、性别及死因无关(P>0.05)。结论 U6可以作为实时RT-PCR方法研究死亡时间与心肌组织中核酸降解之间规律的理想内对照。Objective To explore the stability of internal controls in human cardiac muscle by real-time RT-PCR during early postmortem interval（PMI） in order to find the most stable marker.Methods Ten individuals with similar environmental conditions（the average store temperature： 25 ℃） and different PMI ranging from 4.3 to 22.3 h were selected.Total RNA was extracted from each sample and six commonly internal controls were used including β-actin,GAPDH,B2M,U6,18S rRNA and HSA-miR-1,and the expression was detected in cardiac muscle by real-time RT-PCR.The expression stability of internal controls was evaluated using genormPLUS software during early PMI.The internal control with the most stability was selected.The relationship between the most stable marker and its expression level affected by some other parameters such as age,gender and cause of death was also analyzed.Results The U6 showed the most stable expression during early PMI in cardiac muscle,and its expression level was not affected by those parameters including age,gender and cause of death（P0.05）.Conclusion U6 may be a valuable internal control for the study of relationship between PMI determination and degradation of nucleic acid in human cardiac muscle by real-time RT-PCR.

关 键 词：法医病理学 逆转录聚合酶链反应 心肌 内对照 死亡时间 

分 类 号：R735.2[医药卫生—肿瘤]