Rapid Identification of Plasma DNA Samples with Increased ctDNA Levels by a Modified FAST-SeqS Approach
| Autor: | Thomas Bauernhofer, Marina Koch, Martina Auer, Katja Fischereder, Jochen B. Geigl, Jelena Belic, Sumitra Mohan, Teresa Gerhalter, Michael R. Speicher, Peter Ulz, Ellen Heitzer, Edgar Petru |
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| Rok vydání: | 2015 |
| Předmět: |
Adult
Male Sequence analysis Clinical Biochemistry Mutant Aneuploidy Biology Sensitivity and Specificity Genome chemistry.chemical_compound Reference Values Neoplasms medicine Humans Allele Allele frequency Aged Aged 80 and over Biochemistry (medical) High-Throughput Nucleotide Sequencing Prostatic Neoplasms Reproducibility of Results Chromosome DNA Sequence Analysis DNA Middle Aged Neoplastic Cells Circulating medicine.disease Molecular biology Genetic Techniques chemistry Case-Control Studies Female |
| Zdroj: | Clinical Chemistry. 61:838-849 |
| ISSN: | 1530-8561 0009-9147 |
| DOI: | 10.1373/clinchem.2014.234286 |
| Popis: | BACKGROUND Recent progress in the analysis of cell-free DNA fragments [cell-free circulating tumor DNA (ctDNA)] now allows monitoring of tumor genomes by noninvasive means. However, previous studies with plasma DNA from patients with cancer demonstrated highly variable allele frequencies of ctDNA. The comprehensive analysis of tumor genomes is greatly facilitated when plasma DNA has increased amounts of ctDNA. Therefore, a fast and cost-effective prescreening method to identify such plasma samples without previous knowledge about alterations in the respective tumor genome could assist in the selection of samples suitable for further extensive qualitative analysis. METHODS We adapted the recently described Fast Aneuploidy Screening Test-Sequencing System (FAST-SeqS) method, which was originally established as a simple, effective, noninvasive screening method for fetal aneuploidy from maternal blood. RESULTS We show that our modified FAST-SeqS method (mFAST-SeqS) can be used as a prescreening tool for an estimation of ctDNA percentage. With a combined evaluation of genome-wide and chromosome arm–specific z-scores from dilution series with cell line DNA and by comparisons of plasma-Seq profiles with data from mFAST-SeqS, we established a detection limit of ≥10% mutant alleles. Plasma samples with an mFAST-SeqS z-score >5 showed results that were highly concordant with those of copy number profiles obtained from our previously described plasma-Seq approach. CONCLUSIONS Advantages of this approach include the speed and cost-effectiveness of the assay and that no prior knowledge about the genetic composition of tumor samples is necessary to identify plasma DNA samples with >10% ctDNA content. |
| Databáze: | OpenAIRE |
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