Abstract
Conventional methods for the isolation of cancer-related circulating cell-free (ccf) DNA from patient blood (plasma) are time consuming and laborious. A DEP approach utilizing a microarray device now allows rapid isolation of ccf-DNA directly from a small volume of unprocessed blood. In this study, the DEP device is used to compare the ccf-DNA isolated directly from whole blood and plasma from 11 chronic lymphocytic leukemia (CLL) patients and one normal individual. Ccf-DNA from both blood and plasma samples was separated into DEP high-field regions, after which cells (blood), proteins, and other biomolecules were removed by a fluidic wash. The concentrated ccf-DNA was detected on-chip by fluorescence, and then eluted for PCR and DNA sequencing. The complete process from blood to PCR required less than 10 min; an additional 15 min was required to obtain plasma from whole blood. Ccf-DNA from the equivalent of 5 μL of CLL blood and 5 μL of plasma was amplified by PCR using Ig heavy-chain variable (IGHV) specific primers to identify the unique IGHV gene expressed by the leukemic B-cell clone. The PCR and DNA sequencing results obtained by DEP from all 11 CLL blood samples and from 8 of the 11 CLL plasma samples were exactly comparable to the DNA sequencing results obtained from genomic DNA isolated from CLL patient leukemic B cells (gold standard).
Original language | English (US) |
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Pages (from-to) | 1828-1836 |
Number of pages | 9 |
Journal | ELECTROPHORESIS |
Volume | 35 |
Issue number | 12-13 |
DOIs | |
State | Published - Jul 2014 |
Externally published | Yes |
Keywords
- Biomarkers
- Cancer
- Chronic lymphocytic leukemia (CLL)
- Circulating cell-free (ccf)-DNA
- Dielectrophoresis
ASJC Scopus subject areas
- Biochemistry
- Clinical Biochemistry