Validation of reference genes for estimating wound age in contused rat skeletal muscle by quantitative real-time PCR
Validation of reference genes for estimating wound age in contused rat skeletal muscle by quantitative real-time PCR
复制标题
通过定量实时 PCR 验证用于估计挫伤大鼠骨骼肌伤口年龄的参考基因
DOI：
10.1007/s00414-011-0604-3
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发表时间：
2011
影响因子：
2.1
通讯作者：
Wang, Ying-yuan
中科院分区：
文献类型：
--
作者：
Sun, Jun-hong;Nan, Liang-huan;Gao, Cai-rong;Wang, Ying-yuan
实时荧光定量PCR （RT-qPCR）基因表达谱是法医鉴定伤口年龄的重要工具。为了准确评估受损组织中基因随时间的表达水平，必须仔细验证用作内部参考标准的基因的转录稳定性。本研究通过RT-qPCR检测了挫伤大鼠骨骼肌中9个潜在内参基因（β-actin、GAPDH、RPL32、PGK1、SDHA、RPL13、HPRT、Tbp和Ywhaz）的转录稳定性。在挫伤后的不同时间点测定每个候选基因的原始Ct值，并通过NormFinder、geNorm和BestKeeper验证程序分析数据。内参基因RPL13和RPL32在挫伤骨骼肌中表达最稳定，而PGK1表达最不稳定。常用的内参基因β-肌动蛋白和GAPDH似乎太不稳定，无法规范损伤肌肉的RT-qPCR表达谱。内参基因RPL13和RPL32也为多重分析的最佳组合。使用RPL13和RPL32作为内部标准可以提高基因表达研究的准确性，旨在确定法医调查中早期伤口的年龄。
Gene expression profiling by quantitative real-time PCR (RT-qPCR) is a valuable tool in forensic science for estimating the age of a wound. To accurately assess gene expression levels over time in injured tissue, the genes used as internal reference standards must be carefully validated for transcriptional stability. This study examined the transcriptional stability of nine potential reference genes (β-actin, GAPDH, RPL32, PGK1, SDHA, RPL13, HPRT, Tbp, and Ywhaz) in contused rat skeletal muscle by RT-qPCR. The raw Ct values were determined for each candidate gene at different time points following contusion, and the data were analyzed by the NormFinder, geNorm, and BestKeeper validation programs. The reference genes RPL13 and RPL32 were the most stably expressed genes in contused skeletal muscle, whereas PGK1 was the least stable. The commonly used reference genes β-actin and GAPDH appeared to be too unstable for normalization of RT-qPCR expression profiling in contused muscle. The reference genes RPL13 and RPL32 were also the best combination for multianalysis. The use of RPL13 and RPL32 as internal standards may improve the accuracy of gene expression studies aimed at determining the age of early wounds in forensic investigations.
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影响因子：
3.1
作者：
Young NJ;Thomas CJ;Collins ME;Brownlie J
通讯作者：
Brownlie J
影响因子：
3.3
作者：
Selvey, S;Thompson, EW;Griffiths, LR
通讯作者：
Griffiths, LR
影响因子：
5.4
作者：
Toshihisa Kobayashi;Y. Sasaki;Yuichiro Oshima;Hiroyuki Yamamoto;H. Mita;Hiromu Suzuki;M. Toyota
通讯作者：
Toshihisa Kobayashi;Y. Sasaki;Yuichiro Oshima;Hiroyuki Yamamoto;H. Mita;Hiromu Suzuki;M. Toyota
影响因子：
4.3
作者：
N. McBrier;J. M. Lekan;L. Druhan;S. Devor;M. Merrick
通讯作者：
N. McBrier;J. M. Lekan;L. Druhan;S. Devor;M. Merrick
影响因子：
--
作者：
Curtis KM;Gomez LA;Rios C;Garbayo E;Raval AP;Perez-Pinzon MA;Schiller PC
通讯作者：
Schiller PC