荧光标记RT-PCR技术在实验动物死亡时间推断中的应用

Preliminary study on fluorimetric RT-PCR in estimating postmortem interval in rats

目的探讨荧光标记逆转录-聚合酶联反应(RT-PCR)在死亡时间(PM I)推断中的应用。方法44只大鼠断颈处死后置于20℃温度控制系统内,利用荧光标记RT-PCR技术检测大鼠脑及肝组织GAPDH mRNA在死后即刻至8 d的相对表达量的变化。结果在大鼠死后即刻至7 d脑组织、2 d肝组织内均可检测到GAPDH mRNA,其扩增产物的降解呈现出一定的规律性。结论荧光RT-PCR技术检测大鼠死后脑及肝GAPDH mRNA,可为PM I推断提供一种新的方法。

Objective To investigate the application of fluorimetric reverse transcriptase-polymerase chain reaction（RT-PCR） to estimate postmortem interval（PMI） in rats. Methods A total of 44 rats were killed by cervical dislocation and placed in a chamber with temperature-controlling system at 20℃ for 0 to 8 d postmortem. Changes of the relative content of glyceraldehyde phosphate dehydrogenase（GAPDH） mRNA in the brain and liver were examined by fluofimetfic RT-PCR. Results The amplification of GAPDH mRNA could be detected within 7 d in the brain and 2 d in the liver postmortem respectively,in a fashion of decreasing. Conclusion Fluorimetric RT-PCR can detect the time-dependent degradation of GAPDH mRNA in the brain and the liver, so it maybe provide a new index for estimating PMI.