Dielectrophoretic recovery of DNA from plasma for the identification of chronic lymphocytic leukemia point mutations. (May 2016)

• Record Type: Journal Article
• Authors: Manouchehri, Sareh; Ibsen, Stuart; Wright, Jennifer; Rassenti, Laura; Ghia, Emanuela M; Widhopf, George F; Kipps, Thomas J; Heller, Michael J
• Is Part Of: International journal of hematologic oncology. Volume 5:Number 1(2016)
• Page Start: 27
• Page End: 35
• Publication Date: 2016-05
• Subjects: biomarkers -- chronic lymphocytic leukemia -- circulating cell free DNA -- dielectrophoresis -- electrokinetic -- genetics -- point mutations; Hematological oncology -- Periodicals; 616.99418
• DOI: 10.2217/ijh-2015-0009
• Languages: English
• ISSNs: 2045-1393

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.