Sareh Manouchehri, Stuart Ibsen, Jennifer Wright, Laura Rassenti, Emanuela M Ghia, George F Widhopf, Thomas J Kipps, Michael J Heller
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Dielectrophoretic recovery of DNA from plasma for the identification of chronic lymphocytic leukemia point mutations.
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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