Effects of storage method on DNA degradation in old bloodstain samples
Effects of storage method on DNA degradation in old bloodstain samples
复制标题
DOI：
10.1016/j.fsigss.2015.09.016
复制
发表时间：
2015-12
影响因子：
--
通讯作者：
M. Hara;H. Nakanishi;S. Takahashi;A. Tamura;K. Yoneyama;Kazuyuki Saito;A. Takada
中科院分区：
文献类型：
--
作者：
M. Hara;H. Nakanishi;S. Takahashi;A. Tamura;K. Yoneyama;Kazuyuki Saito;A. Takada
评估了不同储存方法对 DNA 降解和初步血液测试结果的影响。血迹在室温、4°C、–20°C 和–80°C 下保存 20 年；全血样本在 –20 °C 和 –80 °C 下保存 20 年；并对新鲜全血样本进行分析。通过量化 305 和 129 碱基对 (bp) 片段与 41 bp 片段的比率来评估 DNA 降解。初步测试包括无色孔雀石绿染色、通过免疫层析法进行抗人血红蛋白 (Hb) 测试以及血红蛋白-β (HBB) mRNA 的检测。 129:41 bp DNA 片段和 305:41 bp DNA 片段的比例表明，在室温和 4 °C 下保存的血迹比新鲜血液的降解程度更高。使用无色孔雀石绿染色和抗人 Hb 检测，所有样本均呈阳性。尽管在所有血迹样本中均检测到了六溴代苯，但在 –20°C 或 –80°C 下储存的任何全血样本中均未检测到六溴代苯。因此，为防止长期储存过程中 DNA 降解，建议将血液样本储存在 –20 °C 以下。另外，储存的血迹适合用于血液的初步检测，而不是储存血液。
The effects of different storage methods on both DNA degradation and the results of preliminary blood tests were evaluated. Bloodstains stored at room temperature, 4 °C, –20 °C, and –80 °C for 20 years; whole blood samples stored at –20 °C and –80 °C for 20 years; and fresh whole blood samples were analyzed. DNA degradation was evaluated by quantifying the ratios of 305 and 129 base pair (bp) fragments to 41 bp fragments. Preliminary testing included leuco-malachite-green staining, anti-human hemoglobin (Hb) testing by immunochromatography, and the detection of the hemoglobin-beta (HBB) mRNA. Bloodstains stored at room temperature and 4 °C were more highly degraded than fresh blood, as indicated by the ratio of 129:41 bp DNA fragments and 305:41 bp DNA fragments. All samples tested positive using leuco-malachite-green staining and anti-human Hb assays. HBB was not detected in any whole blood samples stored at –20 °C or –80 °C, although HBB was detected in all bloodstain samples. Therefore, to prevent DNA degradation during long-term storage, it is recommended that blood samples be stored at below –20 °C. In addition, stored bloodstains are suitable for preliminary tests of blood, rather than storing blood.