Academic Journal
Assessment of Pre-Analytical Sample Handling Conditions for Comprehensive Liquid Biopsy Analysis
| Title: | Assessment of Pre-Analytical Sample Handling Conditions for Comprehensive Liquid Biopsy Analysis |
|---|---|
| Authors: | Teresa Gerber, Sabine Taschner-Mandl, Lisa Saloberger-Sindhöringer, Niko Popitsch, Ellen Heitzer, Volker Witt, René Geyeregger, Caroline Hutter, Raphaela Schwentner, Inge M. Ambros, Peter F. Ambros |
| Source: | The Journal of Molecular Diagnostics. 22:1070-1086 |
| Publisher Information: | Elsevier BV, 2020. |
| Publication Year: | 2020 |
| Subject Terms: | Adult, Male, 0301 basic medicine, Time Factors, Adolescent, Blood Donors, Circulating Tumor DNA, Young Adult, 03 medical and health sciences, 0302 clinical medicine, 106023 Molekularbiologie, Blood Specimen Collection/methods, Diagnostic Tests, Whole Genome Sequencing/methods, Leukocytes, Humans, Genetic Testing, Edetic Acid, Aged, Blood Specimen Collection, Circulating Tumor DNA/genetics, Molecular Diagnostic Techniques/methods, Diagnostic Tests, Routine, Heparin, Temperature, Liquid Biopsy, 106023 Molecular biology, Liquid Biopsy/methods, Middle Aged, Flow Cytometry, Flow Cytometry/methods, 3. Good health, Heparin/chemistry, Phenotype, Molecular Diagnostic Techniques, Routine/methods, Feasibility Studies, Female, Edetic Acid/chemistry, Genetic Testing/methods |
| Description: | Liquid biopsies as a minimally invasive approach have the potential to revolutionize molecular diagnostics. Yet, although protocols for sample handling and the isolation of circulating tumor DNA (ctDNA) are numerous, comprehensive guidelines for diagnostics and research considering all aspects of real-life multicenter clinical studies are currently not available. These include limitations in sample volume, transport, and blood collection tubes. We tested the impact of commonly used (EDTA and heparin) and specialized blood collection tubes and storage conditions on the yield and purity of cell-free DNA for the application in down-stream analysis. Moreover, we evaluated the feasibility of a combined workflow for ctDNA and tumor cell genomic testing and parallel flow cytometric analysis of leukocytes. For genomic analyses, EDTA tubes showed good results if stored for a maximum of 4 hours at room temperature or for up to 24 hours when stored at 4°C. Spike-in experiments revealed that EDTA tubes in combination with density gradient centrifugation allowed the parallel isolation of ctDNA, leukocytes, and low amounts of tumor cells (0.1%) and their immunophenotyping by flow cytometry and down-stream genomic analysis by whole genome sequencing. In conclusion, adhering to time and temperature limits allows the use of routine EDTA blood samples for liquid biopsy analyses. We further provide a workflow enabling the parallel analysis of cell-free and cellular features for disease monitoring and for clonal evolution studies. |
| Document Type: | Article |
| Language: | English |
| ISSN: | 1525-1578 |
| DOI: | 10.1016/j.jmoldx.2020.05.006 |
| Access URL: | http://www.jmdjournal.org/article/S1525157820303512/pdf https://pubmed.ncbi.nlm.nih.gov/32497717 https://ucrisportal.univie.ac.at/de/publications/88ebad8a-102b-4470-b3fe-6819581492b9 https://doi.org/10.1016/j.jmoldx.2020.05.006 https://pubmed.ncbi.nlm.nih.gov/32497717/ https://www.ncbi.nlm.nih.gov/pubmed/32497717 https://jmd.amjpathol.org/article/S1525-1578(20)30351-2/fulltext https://www.jmdjournal.org/article/S1525-1578(20)30351-2/fulltext https://www.sciencedirect.com/science/article/pii/S1525157820303512 https://www.sciencedirect.com/science/article/abs/pii/S1525157820303512 |
| Rights: | CC BY NC ND |
| Accession Number: | edsair.doi.dedup.....705a1d3783df9c62c3f4da8cf5c4f498 |
| Database: | OpenAIRE |
| ISSN: | 15251578 |
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| DOI: | 10.1016/j.jmoldx.2020.05.006 |