Streck cfDNA采血管液體活檢應(yīng)用表現(xiàn)

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

液體活檢的廣泛應(yīng)用對室溫運(yùn)送血液樣本提出了更高要求，采血管需要保持游離DNA的數(shù)量和穩(wěn)定，防止循環(huán)腫瘤DNA被基因組DNA稀釋。通過高靈敏度突變檢測技術(shù)評(píng)估Streck Cell-Free DNA BCT采血管對細(xì)胞和DNA穩(wěn)定性的保護(hù)。

方法

靜脈血樣分別用Streck Cell-Free DNA BCT和標(biāo)準(zhǔn)K2EDTA管采集。提取DNA之前采血管分別在不同溫度儲(chǔ)存。提取后的游離DNA進(jìn)行qPCR，BEAMing和 Plasma Safe-Sequencing (Safe-SeqS)分析。

結(jié)果

用Streck Cell-Free DNA BCT采集的健康血樣室溫存儲(chǔ)5天后，并不影響DNA產(chǎn)量和突變水平。

結(jié)論

使用Streck Cell-Free DNA BCT采集運(yùn)送的全血可以用于下游液體活檢（BEAMing和 Safe-SeqS）。由于運(yùn)送溫度是重要影響因素，要獲得可靠的突變結(jié)果需要特別注意保持室溫在限定范圍內(nèi)。

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 第二代升級(jí)新型混合塑料采血管

Streck 230253 9mL Hybrid Plastic Cell-Free DNA BCT 第二代升級(jí)新型混合塑料采血管是一種特制混合塑料的新型采血管。特制保存劑能穩(wěn)定有核血細(xì)胞，防止細(xì)胞基因組DNA的釋放，保留血漿游離循環(huán)DNA和循環(huán)腫瘤細(xì)胞（CTC），保證游離DNA的高品質(zhì)分離。

產(chǎn)品訂購信息-Streck 230253 9mL Hybrid Plastic Cell-Free DNA BCT 第二代升級(jí)新型混合塑料采血管

RUO：Research Use Only

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華雅干細(xì)胞 現(xiàn)貨供應(yīng)：美國Streck 218962 Cell-Free DNA BCT Blood Collection Tube DNA基因檢測無創(chuàng)采血管和美國Streck 218976 Cell-Free RNA BCT Blood Collection Tube基因檢測無創(chuàng)采血管 規(guī)格：100管/盒，應(yīng)用于：唐氏無創(chuàng)篩查采血管，游離DNA采血管，游離RNA采血管等等。華雅干細(xì)胞 現(xiàn)貨供應(yīng)：BD 362761 Vacutainer® CPT™ Cell Preparation Tube 肝素鈉單核細(xì)胞制備管/采血管，60支/盒。

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