全血标本保存条件对人总RNA提取效率的影响

RNA isolation from human blood: impacts of sample-storage condition

目的探讨全血标本保存条件对RNA提取效率和RNA完整性的影响。方法从179例保存在不同温度、不同时间的全血标本中提取总RNA,测定其浓度和纯度、分析其完整性。另选20例标本研究冻融次数对RNA提取效率的影响。179例中的73例总RNA在-80℃下保存1周前、后分别测定浓度和纯度,观察两者差异。结果新鲜血来源的总RNA浓度最高,平均306.51ng/μl;冻融2次全血仍可获得足量的RNA,浓度181.98ng/μl,RNA完整性有所下降;-80℃保存1周的RNA,浓度和纯度都略有下降。结论利用Trizol法可从小体积全血标本中获取足量有效的总RNA;不同保存条件全血中所获取的总RNA浓度存在差异;总RNA-80℃短期保存也会略有降解。

Objective To study the impacts of different sample storage conditions. on RNA isolation efficiency and RNA integrity. Methods Total RNA was isolated from peripheral blood samples （179） stored at different temperatures with different durations. Twenty blood samples were used to investigate the impacts from sample freezing and thawing. Total RNA concentration, purity and integrity were tested. Seventy-three of 179 RNA samples were re-tested, after stored at-80℃ for 1 week, with the data compared to the primarily tested results. Results RNA concentration from fresh blood was the highest 306.51ng/μl on average. Samples after two cycles of freeze/thaw were still available for sufficient RNA isolation, with an average concentration of 181.98 ng/μl, but the integrity of RNA decreased slightly. Both the concentration and purity of RNA declined slightly after 1-week storage at-80℃. Conclusions Blood sample storage condition has significant impacts on RNA isolated concentration. Trizol method is quite stable to isolate sufficient amount RNA from small volume of blood. RNA sample degenerates slightly at-80℃ even for a short time.