Streck cfDNA采血管液体活检应用表现

Performance of Streck cfDNA Blood Collection Tubes for Liquid Biopsy Testing

Inga Medina Diaz, Annette Nocon, Daniel H. Mehnert, Johannes Fredebohm, Frank Diehl , Frank Holtrup

Published: November 10, 2016https://doi.org/10.1371/journal.pone.0166354

摘要

目的

液体活检的广泛应用对室温运送血液样本提出了更高要求，采血管需要保持游离DNA的数量和稳定，防止循环肿瘤DNA被基因组DNA稀释。通过高灵敏度突变检测技术评估Streck Cell-Free DNA BCT采血管对细胞和DNA稳定性的保护。

方法

静脉血样分别用Streck Cell-Free DNA BCT和标准K2EDTA管采集。提取DNA之前采血管分别在不同温度储存。提取后的游离DNA进行qPCR，BEAMing和 Plasma Safe-Sequencing (Safe-SeqS)分析。

结果

用Streck Cell-Free DNA BCT采集的健康血样室温存储5天后，并不影响DNA产量和突变水平。

结论

使用Streck Cell-Free DNA BCT采集运送的全血可以用于下游液体活检（BEAMing和 Safe-SeqS）。由于运送温度是重要影响因素，要获得可靠的突变结果需要特别注意保持室温在限定范围内。

Objectives

Making liquid biopsy testing widely available requires a concept to ship whole blood at ambient temperatures while retaining the integrity of the cell-free DNA (cfDNA) population and stability of blood cells to prevent dilution of circulating tumor DNA (ctDNA) with wild-type genomic DNA. The cell- and DNA-stabilizing properties of Streck Cell-Free DNA BCT blood collection tubes (cfDNA BCTs) were evaluated to determine if they can be utilized in combination with highly sensitive mutation detection technologies.

Methods

Venous blood from healthy donors or patients with advanced colorectal cancer (CRC) was collected in cfDNA BCTs and standard K2EDTA tubes. Tubes were stored at different temperatures for various times before plasma preparation and DNA extraction. The isolated cfDNA was analyzed for overall DNA yield of short and long DNA fragments using qPCR as well as for mutational changes using BEAMing and Plasma Safe-Sequencing (Safe-SeqS).

Results

Collection of whole blood from healthy individuals in cfDNA BCTs and storage for up to 5 days at room temperature did not affect the DNA yield and mutation background levels (n = 60). Low-frequency mutant DNA spiked into normal blood samples as well as mutant circulating tumor DNA in blood samples from CRC patients collected in cfDNA BCTs were reliably detected after 3 days of storage at room temperature. However, blood samples stored at ≤ 10°C and at 40°C for an extended period of time showed elevated normal genomic DNA levels and an abnormally large cellular plasma interface as well as lower plasma volumes.

Conclusion

Whole blood shipped in cfDNA BCTs over several days can be used for downstream liquid biopsy testing using BEAMing and Safe-SeqS. Since the shipping temperature is a critical factor, special care has to be taken to maintain a defined room temperature range to obtain reliable mutation testing results.

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Streck 230253 9mL Hybrid Plastic Cell-Free DNA BCT 第二代升级新型混合塑料采血管

Streck 230253 9mL Hybrid Plastic Cell-Free DNA BCT 第二代升级新型混合塑料采血管是一种特制混合塑料的新型采血管。特制保存剂能稳定有核血细胞，防止细胞基因组DNA的释放，保留血浆游离循环DNA和循环肿瘤细胞（CTC），保证游离DNA的高品质分离。

产品订购信息-Streck 230253 9mL Hybrid Plastic Cell-Free DNA BCT 第二代升级新型混合塑料采血管

RUO：Research Use Only

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Streck 230253 9mL Hybrid Plastic Cell-Free DNA BCT 第二代升级新型混合塑料采血管，Streck Cell-Free DNA BCT Blood Collection Tube 218976, 218962基因无创管，唐氏无创筛查采血管，游离DNA采血管，游离RNA采血管，循环肿瘤细胞（circulating tumor cell）CTC采血管 华雅再生医学 现货供应

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