荧光半定量逆转录-聚合酶链式反应检测大鼠死后肾mRNA

Detection of mRNA in kidney of dead rat using fluorescence semi-quantitative RT-PCR

目的探讨荧光半定量逆转录-聚合酶链式反应(fluorescencesemi-quantitativereversetranscrip-tase-polymerasechainreaction,RT-PCR)技术检测大鼠死后不同时间肾3-磷酸甘油醛脱氢酶(GAPDH)mRNA含量变化对死亡时间(postmorteminterval,PMI)推断的应用价值。方法28只大鼠断颈处死后置于20℃温控系统内,利用荧光标记半定量RT-PCR技术检测大鼠肾GAPDHmRNA在死后48h内相对表达量的变化情况。结果在死后即刻至40h内大鼠肾均可检测出GAPDHmRNA,且其扩增产物呈规律性下降。结论荧光半定量RT-PCR技术检测大鼠死后肾GAPDHmRNA含量变化对PMI推断具有一定的应用价值。

[Objective ] To explore the value of fluorescence semi-quantitatlve reverse transcriptase-polymerase chain reaction （RT-PCR） technique in detecting Glyceraldehyde phosphate dehydrogenase （GAPDH） mRNA in the kidney of rat in estimation of postmortem intervals （PMI）. [Methods] 28 rats were killed by cervical dislocation and placed in a chamber with temperature-controlling system at 20℃ for 48 h postmortem. Changes of GAPDH mRNA in kidney were examined by fluorescence semi-quantitative RT-PCR. [Results] The amplification products of GAPDH mRNA could be detected in kidney within 40 h postmortem, which showed a decreasing tendency. [Conclusion] Time-dependent degradation of GAPDH mRNA in kidney as detected by fluorescence semi-quantitative RT-PCR technique has some value for estimation of PMI.