Streck, Inc. Omaha, NE 68128, USA.
J Clin Lab Anal. 2013 Jul;27(4):305-11. doi: 10.1002/jcla.21603.
Cell-free DNA (cfDNA) circulating in blood is currently used for noninvasive diagnostic and prognostic tests. Minimizing background DNA is vital for detection of low abundance cfDNA. We investigated whether a new blood collection device could reduce background levels of genomic DNA (gDNA) in plasma compared to K(3) EDTA tubes, when subjected to conditions that may occur during sample storage and shipping.
Blood samples were drawn from healthy donors into K(3) EDTA and Cell-Free DNA™ BCT (BCT). To simulate shipping, samples were shaken or left unshaken. In a shipping study, samples were shipped or not shipped. To assess temperature variations, samples were incubated at 6°C, 22°C, and 37°C. In all cases, plasma was harvested by centrifugation and total plasma DNA (pDNA) assayed by quantitative real-time polymerase chain reaction (qPCR).
Shaking and shipping blood in K(3) EDTA tubes showed significant increases in pDNA, whereas no change was seen in BCTs. Blood in K(3) EDTA tubes incubated at 6°C, 22°C, and 37°C showed increases in pDNA while pDNA from BCTs remained stable.
BCTs prevent increases in gDNA levels that can occur during sample storage and shipping. This new device permits low abundance DNA target detection and allows accurate cfDNA concentrations.
目前,游离于血液中的细胞外 DNA(cfDNA)被用于非侵入性诊断和预后检测。为了检测低丰度 cfDNA,减少背景 DNA 至关重要。本研究旨在比较 K3 EDTA 管和 Cell-Free DNA™ BCT(BCT)在储存和运输过程中可能出现的条件下,新的采血装置是否能降低血浆中基因组 DNA(gDNA)的背景水平。
从健康供体中抽取血液至 K3 EDTA 和 Cell-Free DNA™ BCT(BCT)中。为了模拟运输条件,对样品进行了摇匀或不摇匀处理。在运输研究中,对样品进行了运输或不运输处理。为了评估温度变化,将样品孵育在 6°C、22°C 和 37°C 下。在所有情况下,通过离心收集血浆并通过实时定量聚合酶链反应(qPCR)检测总血浆 DNA(pDNA)。
K3 EDTA 管中血液的摇匀和运输导致 pDNA 显著增加,而 BCT 中未见变化。在 6°C、22°C 和 37°C 孵育的 K3 EDTA 管中的血液中 pDNA 增加,而 BCT 中的 pDNA 保持稳定。
BCT 可防止储存和运输过程中 gDNA 水平的升高。这种新装置允许检测低丰度 DNA 靶标,并可实现准确的 cfDNA 浓度检测。