Liquid biopsy - Performance of the PAXgene® Blood ccfDNA Tubes for the isolation and characterization of cell-free plasma DNA from tumor patients
| Autor: | Michael Fleischhacker, Ines Bork, Iris Reindl, Bernd Schmidt, Dana Reinicke, Bettina Wollschläger, Nina Lambrecht |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Lung Neoplasms Lysis Clinical Biochemistry Cell free Biology Biochemistry Circulating Tumor DNA Extended storage 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Humans Liquid biopsy Blood Specimen Collection Plasma dna Biochemistry (medical) Liquid Biopsy General Medicine Isolation (microbiology) DNA extraction Molecular biology 030104 developmental biology chemistry 030220 oncology & carcinogenesis DNA |
| Zdroj: | Clinica Chimica Acta. 469:94-98 |
| ISSN: | 0009-8981 |
| Popis: | Background In most research laboratories the use of EDTA tubes for the isolation of plasma DNA from tumor patients is standard. Unfortunately these tubes do not allow for an extended storage of samples before processing and prevent EDTA tubes from being shipped at ambient temperature. The aim of our study was to compare the quantity and quality of plasma DNA isolated from EDTA and PAXgene® Blood ccfDNA Tubes in different downstream applications. Methods We enrolled 29 patients in our study. Blood samples were drawn into EDTA and PAXgene® Blood ccfDNA Tubes and were processed on day 0 and day 7 after storage at ambient temperature. The plasma DNA from 10 patients was isolated manually. For the DNA isolation from the plasma of 19 additional patients we used the automated QIAsymphony system. The total amount DNA from all samples was measured with a quantitative real-time PCR assay. In addition the amount of methylated mSHOX2 plasma DNA was determined. Results While the 7 day storage lead to an increased amount of total DNA in almost all EDTA tubes, this effect was only seen in very few PAXgene® Blood ccfDNA Tubes. The stabilization solution which prevents the lysis of blood cells had no effect on the method for quantification of methylated sequences in these samples. Conclusion The quantity and quality of plasma DNA from both types of blood draw tubes are comparable. DNA from PAXgene® Blood ccfDNA was successfully used for PCR-based quantification of total amount of cell-free DNA and for methylation analysis as well. |
| Databáze: | OpenAIRE |
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