Plasma vs. serum in circulating tumor DNA measurement: characterization by DNA fragment sizing and digital droplet polymerase chain reaction
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Jee-Soo Lee
Abstract
Background
Choosing the specimen type is the first step of the pre-analytical process. Previous reports suggested plasma as the optimal specimen for circulating tumor DNA (ctDNA) analysis. However, head-to-head comparisons between plasma and serum using platforms with high analytical sensitivity, such as droplet digital polymerase chain reaction (ddPCR), are limited, and several recent studies have supported the clinical utility of serum-derived ctDNA. This study aimed to compare the DNA profiles isolated from plasma and serum, characterize the effects of the differences between specimens on ctDNA measurement, and determine the major contributors to these differences.
Methods
We isolated cell-free DNA (cfDNA) from 119 matched plasma/serum samples from cancer patients and analyzed the cfDNA profiles by DNA fragment sizing. We then assessed KRAS mutations in ctDNA from matched plasma/serum using ddPCR.
Results
The amount of large DNA fragments was increased in serum, whereas that of cfDNA fragments (<800 bp) was similar in both specimens. ctDNA was less frequently detected in serum, and the KRAS-mutated fraction in serum was significantly lower than that in plasma. The differences in ctDNA fractions between the two specimen types correlated well with the amount of large DNA fragments and white blood cell and neutrophil counts.
Conclusions
Our results provided detailed insights into the differences between plasma and serum using DNA fragment sizing and ddPCR, potentially contributing to ctDNA analysis standardization. Our study also suggested that using plasma minimizes the dilution of tumor-derived DNA and optimizes the sensitivity of ctDNA analysis. So, plasma should be the preferred specimen type.
Funding source: Hallym University
Award Identifier / Grant number: HURF-2018-01
Funding statement: This work was supported by the Hallym University Research Fund (HURF-2018-01).
Author contributions: All authors have accepted responsibility for the entire content of the submitted manuscript and have approved its submission.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
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©2020 Walter de Gruyter GmbH, Berlin/Boston
Artikel in diesem Heft
- Frontmatter
- Editorial
- New insights on the analytical performances for detecting and quantifying monoclonal proteins
- Reviews
- Vitamin C measurement in critical illness: challenges, methodologies and quality improvements
- Early predictors of perinatal brain damage: the role of neurobiomarkers
- Opinion Paper – Point
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- EFLM Consensus Paper
- Quantifying atherogenic lipoproteins for lipid-lowering strategies: consensus-based recommendations from EAS and EFLM
- Guidelines and Recommendations
- PREDICT: a checklist for preventing preanalytical diagnostic errors in clinical trials
- Genetics and Molecular Diagnostics
- Plasma vs. serum in circulating tumor DNA measurement: characterization by DNA fragment sizing and digital droplet polymerase chain reaction
- General Clinical Chemistry and Laboratory Medicine
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- Reference Values and Biological Variations
- Influence of ethnicity on biochemical markers of health and disease in the CALIPER cohort of healthy children and adolescents
- Cardiovascular Diseases
- Clinical evaluation of capillary B-type natriuretic peptide testing
- Infectious Diseases
- Thrombo-inflammatory prognostic score improves qSOFA for risk stratification in patients with sepsis: a retrospective cohort study
- Corrigendum
- Corrigendum to: Pediatric reference intervals for 29 Ortho VITROS 5600 immunoassays using the CALIPER cohort of healthy children and adolescents
- Letters to the Editor
- Potential risks in fecal microbiota transplantation
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Artikel in diesem Heft
- Frontmatter
- Editorial
- New insights on the analytical performances for detecting and quantifying monoclonal proteins
- Reviews
- Vitamin C measurement in critical illness: challenges, methodologies and quality improvements
- Early predictors of perinatal brain damage: the role of neurobiomarkers
- Opinion Paper – Point
- Mixing studies for lupus anticoagulant: mostly yes, sometimes no
- Opinion Paper – Counterpoint
- Mixing studies for lupus anticoagulant: mostly no, sometimes yes
- EFLM Consensus Paper
- Quantifying atherogenic lipoproteins for lipid-lowering strategies: consensus-based recommendations from EAS and EFLM
- Guidelines and Recommendations
- PREDICT: a checklist for preventing preanalytical diagnostic errors in clinical trials
- Genetics and Molecular Diagnostics
- Plasma vs. serum in circulating tumor DNA measurement: characterization by DNA fragment sizing and digital droplet polymerase chain reaction
- General Clinical Chemistry and Laboratory Medicine
- An international multi-center serum protein electrophoresis accuracy and M-protein isotyping study. Part I: factors impacting limit of quantitation of serum protein electrophoresis
- An international multi-center serum protein electrophoresis accuracy and M-protein isotyping study. Part II: limit of detection and follow-up of patients with small M-proteins
- Use of clinical data and acceleration profiles to validate pneumatic transportation systems
- National survey on delta checks in clinical laboratories in China
- Assessment of the glomerular filtration rate (GFR) in kidney transplant recipients using Bayesian estimation of the iohexol clearance
- Performance of Afinion HbA1c measurements in general practice as judged by external quality assurance data
- Renal tubular epithelial cells add value in the diagnosis of upper urinary tract pathology
- Reference Values and Biological Variations
- Influence of ethnicity on biochemical markers of health and disease in the CALIPER cohort of healthy children and adolescents
- Cardiovascular Diseases
- Clinical evaluation of capillary B-type natriuretic peptide testing
- Infectious Diseases
- Thrombo-inflammatory prognostic score improves qSOFA for risk stratification in patients with sepsis: a retrospective cohort study
- Corrigendum
- Corrigendum to: Pediatric reference intervals for 29 Ortho VITROS 5600 immunoassays using the CALIPER cohort of healthy children and adolescents
- Letters to the Editor
- Potential risks in fecal microbiota transplantation
- Macro-aspartate aminotransferase syndrome: a case report
- Factitious severe acidosis in a patient, preanalytical considerations and prevention
- Methods for quick, accurate and cost-effective determination of the type 1 diabetes genetic risk score (T1D-GRS)
- Anti-adalimumab and anti-certolizumab antibodies titers after discontinuation of adalimumab: two case reports
- Serum immunoglobulin-A (IgA) concentrations in a general adult population: association with demographics and prevalence of selective IgA deficiency
- Evaluation of PFA-100 closure times in cord blood samples of healthy term and preterm neonates
- Disagreement between direct and indirect potentiometric Na+ determination in infancy and childhood
- Carryover issues with UF-5000 urine flow cytometry – how did we miss it?
