期刊
JOURNAL OF CLINICAL LABORATORY ANALYSIS
卷 27, 期 4, 页码 305-311出版社
WILEY
DOI: 10.1002/jcla.21603
关键词
hematology; investigative techniques; clinical laboratory techniques; blood preservation; real-time polymerase chain reaction
Background: 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. Methods: 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 degrees C, 22 degrees C, and 37 degrees C. In all cases, plasma was harvested by centrifugation and total plasma DNA (pDNA) assayed by quantitative real-time polymerase chain reaction (qPCR). Results: 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 degrees C, 22 degrees C, and 37 degrees C showed increases in pDNA while pDNA from BCTs remained stable. Conclusions: 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. (C) 2013 Wiley Periodicals, Inc.
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