摘要
目的 探究保存剂是否可以达到血液样品在常温储存以及运输中稳定有核细胞,维持血浆中游离DNA丰度的要求.方法 志愿者们分别使用 EDTA-K2和 Vangenes？Cell-Free DNA 采血管各采集一管血液进行常温保存测试,同时使用Streck？Cell-Free DNA和Vangenes？Cell-Free DNA采血管各采集一管血液进行运输保存测试.然后,在特定时间点分离血浆并提取游离DNA,荧光定量检测游离DNA浓度.结果 在室温条件下,血液样品在Vangenes？Cell-Free DNA采血管保存14 d的游离DNA的拷贝数和0 d无显著差异.运输72 h内,Vangenes？Cell-Free DNA 采血管保存血液样品与Streck？Cell-Free DNA采血管效果一致,与0 d拷贝数均无显著差异;游离DNA的PCR扩增效率与0天基本一致.但是,运输7 d,保存剂对游离DNA的PCR扩增存在一定的抑制作用.结论 室温保存中,Vangenes？Cell-Free DNA 采血管可以稳定血液中的有核细胞,维持血浆游离DNA的丰度达到14 d;运输过程中,保存血液样品长达72 h.未来,Vangenes？Cell-Free DNA采血管可用于利用游离DNA丰度变化进行的检测.
Objective To study the ability of a novel stabilizing reagent to prevent nucleated blood cell disruption and genomic DNA contamination during whole blood sample storage and shipping at room temperature（RT）.Methods Samples were drawn from healthy donors into EDTA-K2 and Vangenes Cell-Free DNA BCTs and stored at room temperature or Streck Cell-Free DNA and Vangenes Cell-Free DNA BCTs and shipped round trip.Then,Aliquots were removed at specified time points and cfDNA was purified from the plasma.CfDNA concentration was measured by real-time PCR.Results They indicate that exposure of cfDNA to the Vangenes Cell-Free DNA collection device for up to 14 days had no effect on amplification by PCR as compared to the untreated control samples.In shipping study,blood in Vangenes Cell-Free DNA tubes and Streck Cell-Free DNA tubes had same result after a shipping period of 72 hours,which had no significant differents of copy number and amplification efficiency by PCR.However,there was decrease in DNA amplification in the cfDNA extracted from the plasma separated from blood samples treated by a 7 days ship-ping.Conclusion we concluded that the new reagent could stabilize the nucleated blood cell in whole blood samples and keep con-centration of cfDNA from genomic DNA contamination in plasma after storage at RT up to 14days.In addition,it′s have effect on DNA amplification when blood sample exposed to the reagent for a shipping period of 72 hours.It will replace Streck Cell-Free DNA in downstream analysis of cfDNA in the future.
出处
《检验医学与临床》
CAS
2017年第A02期25-29,共5页
Laboratory Medicine and Clinic
基金
厦门市科技创新基金项目（3502220151086）.
关键词
保存剂
游离DNA
运输
储存
stablizing reagents
cell free DNA
shipping
storage
