Dielectrophoretic recovery of DNA from plasma for the identification of chronic lymphocytic leukemia point mutations.

International Journal of Hematologic Oncology Pub Date : 2016-05-01 Epub Date: 2016-05-05 DOI:10.2217/ijh-2015-0009
Sareh Manouchehri, Stuart Ibsen, Jennifer Wright, Laura Rassenti, Emanuela M Ghia, George F Widhopf, Thomas J Kipps, Michael J Heller
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引用次数: 20

Abstract

Aim: Circulating cell free (ccf) DNA contains information about mutations affecting chronic lymphocytic leukemia (CLL). The complexity of isolating DNA from plasma inhibits the development of point-of-care diagnostics. Here, we introduce an electrokinetic method that enables rapid recovery of DNA from plasma.

Materials & methods: ccf-DNA was isolated from 25 µl of CLL plasma using dielectrophoresis. The DNA was used for PCR amplification, sequencing and analysis.

Results: The ccf-DNA collected from plasma of 5 CLL patients revealed identical mutations to those previously identified by extracting DNA from CLL cells from the same patients.

Conclusion: Rapid dielectrophoresis isolation of ccf-DNA directly from plasma provides sufficient amounts of DNA to use for identification of point mutations in genes associated with CLL progression.

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血浆DNA介电恢复鉴定慢性淋巴细胞白血病点突变。
目的:循环游离细胞(ccf) DNA包含影响慢性淋巴细胞白血病(CLL)的突变信息。从血浆中分离DNA的复杂性抑制了即时诊断技术的发展。在这里,我们介绍了一种能够从血浆中快速恢复DNA的电动方法。材料与方法:从25µl CLL血浆中分离ccf-DNA。将DNA进行PCR扩增、测序和分析。结果:从5例CLL患者的血浆中收集的ccf-DNA显示出与先前从同一患者的CLL细胞中提取DNA所鉴定出的相同的突变。结论:直接从血浆中快速分离ccf-DNA提供了足够数量的DNA用于鉴定与CLL进展相关的基因点突变。
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3
审稿时长
13 weeks
期刊介绍: International Journal of Hematologic Oncology welcomes unsolicited article proposals. Email us today to discuss the suitability of your research and our options for authors, including Accelerated Publication. Find out more about publishing open access with us here.
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