A New Method for Improving Extraction Efficiency and Purity of Urine and Plasma Cell-Free DNA.
A New Method for Improving Extraction Efficiency and Purity of Urine and Plasma Cell-Free DNA.
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一种提高尿液和血浆游离DNA提取效率和纯度的新方法
DOI：
10.3390/diagnostics11040650
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发表时间：
2021-04-03
期刊：
影响因子：
--
通讯作者：
Su YH
中科院分区：
文献类型：
--
作者：
Lin SY;Luo Y;Marshall MM;Johnson BJ;Park SR;Wang Z;Su YH
这项研究评估了三种商用的无细胞DNA(CfDNA)提取试剂盒，以及基于聚乙二醇的DNA清理程序(DNApure)对cfDNA质量和产量的影响。用JBS cfDNA提取试剂盒(KIT J)、MagMAX无细胞DNA提取试剂盒(KIT M)和QIAamp循环核酸试剂盒(KIT Q)对6例正常供者尿液和血浆标本以及4例孕期男性(PG)孕妇的尿液和血浆标本进行提取。回收聚合酶链式反应产物、内源性TP53和Y染色体DNA用于评估试剂盒的性能。核小体大小的DNA图谱在不同的试剂盒中不同，仅通过试剂盒J和M分离的尿液和血浆DNA中存在显著的多核小体大小的峰。试剂盒J比试剂盒M或Q(p<；0.001)从尿液中回收的刺激性dna显著多，从血浆中回收的量相似(p=0.12)。应用DNApure试剂盒可显著提高尿液(p<；0.001)和血浆(p≤0.013)的扩增效率。此外，与试剂盒Q相比，试剂盒J从PG尿液中提取的Y染色体DNA明显多于试剂盒Q(p=0.05)。我们证明，DNApure可以提供一种有效的手段来提高cfDNA的产量和纯度，并将分析前生物样品的变异性对液体活检检测性能的影响降至最低。
This study assessed three commercially available cell-free DNA (cfDNA) extraction kits and the impact of a PEG-based DNA cleanup procedure (DNApure) on cfDNA quality and yield. Six normal donor urine and plasma samples and specimens from four pregnant (PG) women carrying male fetuses underwent extractions with the JBS cfDNA extraction kit (kit J), MagMAX Cell-Free DNA Extraction kit (kit M), and QIAamp Circulating Nucleic Acid Kit (kit Q). Recovery of a PCR product spike-in, endogenous TP53, and Y-chromosome DNA was used to assess kit performance. Nucleosomal-sized DNA profiles varied among the kits, with prominent multi-nucleosomal-sized peaks present in urine and plasma DNA isolated by kits J and M only. Kit J recovered significantly more spike-in DNA than did kits M or Q (p < 0.001) from urine, and similar amounts from plasma (p = 0.12). Applying DNApure to kit M- and Q-isolated DNA significantly improved the amplification efficiency of spike-in DNA from urine (p < 0.001) and plasma (p ≤ 0.013). Furthermore, kit J isolated significantly more Y-chromosome DNA from PG urine compared to kit Q (p = 0.05). We demonstrate that DNApure can provide an efficient means of improving the yield and purity of cfDNA and minimize the effects of pre-analytical biospecimen variability on liquid biopsy assay performance.
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影响因子：
3.7
作者：
Tsui NB;Jiang P;Chow KC;Su X;Leung TY;Sun H;Chan KC;Chiu RW;Lo YM
通讯作者：
Lo YM
影响因子：
8.6
作者：
Gomes, Gabriela A.;Azevedo, Ana M.;Prazeres, D. Miguel F.
通讯作者：
Prazeres, D. Miguel F.
DOI：
10.1073/pnas.1500076112
发表时间：
2015-03-17
影响因子：
11.1
作者：
Jiang, Peiyong;Chan, Carol W. M.;Lo, Y. M. Dennis
通讯作者：
Lo, Y. M. Dennis
影响因子：
4.6
作者：
Markus H;Contente-Cuomo T;Farooq M;Liang WS;Borad MJ;Sivakumar S;Gollins S;Tran NL;Dhruv HD;Berens ME;Bryce A;Sekulic A;Ribas A;Trent JM;LoRusso PM;Murtaza M
通讯作者：
Murtaza M
影响因子：
2.1
作者：
Arbeli, Ziv;Fuentes, Cilia L.
通讯作者：
Fuentes, Cilia L.