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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
【摘要】 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。
【Abstract】 Objective To explore the value of fluorescence semi-quantitative 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.
【Key words】 postmortem interval (PMI); fluorescence semi-quantitative technique; reverse transcriptase-polymerase chain reaction (RT-PCR); glyceraldehyde phosphate dehydrogenase (GAPDH);
- 【文献出处】 中国现代医学杂志 ,China Journal of Modern Medicine , 编辑部邮箱 ,2006年11期
- 【分类号】D919
- 【被引频次】7
- 【下载频次】190