Quantitative analysis of DNA degradation in the dead body.
Quantitative analysis of DNA degradation in the dead body.
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DOI：
10.18926/amo/47011
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发表时间：
2011-06
影响因子：
0.5
通讯作者：
Miki Itani;Yuji Yamamoto;Y. Doi;S. Miyaishi
中科院分区：
文献类型：
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作者：
Miki Itani;Yuji Yamamoto;Y. Doi;S. Miyaishi
定量估计死后DNA的降解。在20℃和4℃下分别取大鼠的脑、肝、肾和肌肉样品。利用Rsrc 1基因序列的引物，采用实时荧光定量PCR测定DNA的数量。当使用10ng人类基因组DNA扩增DNA的数量定义为100 RFU时，祭祀当天脑、肝、肾和骨骼肌（每2μg干重）的数量分别为253±11、338±22、556±14和531±12相对荧光单位（RFU）（平均±S.E.）(n = 5)。在20℃下，肝脏、肾脏和骨骼肌的DNA扩增量在1-3周内下降到10 RFU以下，而大脑的DNA扩增量在6周内超过10 RFU，证明了大脑作为腐烂尸体DNA分析样本的有用性。结果表明，测定20℃时脑内和4℃时肝内DNA扩增量以及4℃时肝内DNA扩增量与脑内DNA扩增量之比，可能有助于诊断死亡时间。这项研究首次提供了尸体DNA降解过程的定量分析。
Postmortem degradation of DNA was quantitatively estimated. Brain, liver, kidney and muscle samples were obtained from sacrificed rats left at 20℃ or 4℃. The quantity of DNA was measured by real-time PCR using a primer set for a sequence in the Rsrc 1 gene. When the quantity of amplified DNA using 10ng Human Genomic DNA was defined as 100 RFU, the quantities in the brain, liver, kidney and skeletal muscle (each 2μg of dry weight) on the day of sacrifice were 253±11, 338±22, 556±14 and 531±12 Relative Fluorescence Units (RFU), respectively (mean±S.E., n＝5). The quantity of amplified DNA decreased to below 10 RFU in 1-3 weeks in the liver, kidney and skeletal muscle at 20℃, while that in the brain was more than 10 RFU for six weeks, demonstrating the usefulness of the brain as a sample for DNA analysis of decaying corpses. It was suggested that quantifying the amplified DNA in the brain at 20℃ and in the liver at 4℃ as well as the ratio of the quantity of amplified DNA in the liver to the brain at 4℃ might be useful for diagnosing time of death. This study provides the first quantitative analysis of the postmortem progress of DNA degradation in the corpse.