Parackal Sarah, Zou Donghui, Day Robert, Black Michael, Guilford Parry
Cancer Genetics Laboratory, Department of Biochemistry, University of Otago, Dunedin New Zealand.
Pract Lab Med. 2019 Jun 18;16:e00125. doi: 10.1016/j.plabm.2019.e00125. eCollection 2019 Aug.
To compare the Roche Cell-Free DNA Collection Tubes against the Streck Cell-Free DNA BCTs for sample stability using Cell Free DNA (cfDNA) from healthy volunteers (n = 20).
DESIGN & METHODS: Whole blood was drawn into five Roche and five Streck tubes per volunteer, stored at room temperature and processed at five different time points (Days 0, 4, 7, 10 and 14). One volunteer had blood drawn into ×10 KEDTA tubes to observe the effect of no preservation buffer on White Blood Cell (WBC) lysis. DNA was extracted from the plasma and the concentration (ng/μL) measured using the Qubit Fluorometer at each time point. The eluted DNA was further analysed by capillary electrophoresis to determine the proportion of cfDNA and gDNA contamination in the samples over the 14 days.
There was no difference in individual (0.097) and median paired (0.26) DNA concentration across the five time points between the two tubes. However, a difference was observed for samples in the Roche tubes for pair days 0-7 ( = 0.01), 0 to 10 (0.046) and 0 to 14 ( = 0.0016) in contrast to the Streck tubes after adjustment for multiple testing.
The findings of this study indicate that the Roche Cell-Free DNA Collection Tubes are a suitable alternative for sample collection and storage at room temperature, albeit for a duration of less than 7 days.
使用来自健康志愿者(n = 20)的游离DNA(cfDNA),比较罗氏游离DNA采集管和Streck游离DNA BCTs在样本稳定性方面的差异。
每位志愿者的全血分别采集到5支罗氏管和5支Streck管中,室温保存,并在5个不同时间点(第0、4、7、10和14天)进行处理。一名志愿者的血液被采集到×10 KEDTA管中,以观察无保存缓冲液对白细胞(WBC)裂解的影响。在每个时间点从血浆中提取DNA,并使用Qubit荧光计测量浓度(ng/μL)。对洗脱的DNA进一步进行毛细管电泳分析,以确定14天内样本中cfDNA和gDNA污染的比例。
在两个管的五个时间点之间,个体DNA浓度(0.097)和配对中位数DNA浓度(0.26)没有差异。然而,在对多重检验进行校正后,与Streck管相比,罗氏管中样本在第0 - 7天(= 0.01)、第0至10天(0.046)和第0至14天(= 0.0016)存在差异。
本研究结果表明,罗氏游离DNA采集管是室温下样本采集和储存的合适替代品,尽管保存时间少于7天。