Study on the admission levels of circulating cell-free DNA in patients with acute myocardial infarction using different quantification methods.
Author(s): Agiannitopoulos K, Samara P, Papadopoulou E, Tsamis K, Mertzanos G, Babalis D, Lamnissou K
Publication: Scand J Clin Lab Invest, 2020, Vol. , Page 1-3
PubMed ID: 32077765 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to compare quantification methods for the measurement of cell-free DNA (cfDNA) levels in plasma from patients suffering ST elevation myocardial infarction and healthy controls. The sensitivity and specificity of cfDNA levels quantified by different methods to diagnose myocardial infarct was also investigated.
Conclusion of Paper
cfDNA quantification differed among the methods and the similarity to quantification by TaqMan real-time PCR was dependent on the presence of myocardial infarct. However, regardless of quantification method, cfDNA concentrations were higher in patients with myocardial infarct than healthy controls and all quantification methods detected myocardial infarct with a sensitivity of 100% and specificity of 91.25% (NanoDrop spectrophotometer) to 98.75% (Qubit 3.0 single-stranded assay or the real-time PCR).
Studies
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Study Purpose
The purpose of this study was to compare quantification methods for the measurement of cfDNA levels in plasma from patients suffering ST elevation myocardial infarction and healthy controls. The sensitivity and specificity of cfDNA levels quantified by different methods to diagnose myocardial infarct was also investigated. Blood was obtained from 80 patients during ST elevation myocardial infarction and 50 healthy controls. Plasma was obtained by an unspecified method and cfDNA was extracted using the QIAamp Circulating Nucleic Acid Kit. cfDNA was quantified using single (ss) and double (ds) stranded Qubit 3.0 DNA Assay Kits, NanoDrop, and real-time PCR based TaqMan Copy Number Reference Assay TERT.
Summary of Findings:
Difference among cfDNA quantification methods were dependent on the presence of myocardial infarct. Quantified levels using the single-stranded Qubit 3.0 assay were only slightly higher than those found by TaqMan real-time PCR in patients with myocardial infarct (48.47 versus 42.83 ng/µL), but measured levels were much higher in healthy controls using the single-stranded Qubit 3.0 assay (8.163 versus 1.178 ng/µL). When quantified using the double-stranded Qubit 3.0 assay, much less cfDNA was quantified than by TaqMan real-time PCR in patients suffering myocardial infarct (6.724 versus 42.83 ng/µL) but the difference was much smaller in healthy controls (1.178 versus 1.420 ng/µL). NanoDrop quantified only half as much cfDNA by TaqMan real-time PCR in those suffering myocardial infarct (22.04 versus 42.83 ng/µL), but much more cfDNA was quantified in healthy controls using NanoDrop (7.771 versus 1.420 ng/µL). As expected, cfDNA concentrations were higher in patients with myocardial infarct than healthy controls, regardless of quantification method (P<0.001, all), and all quantification methods detected myocardial infarct with a sensitivity of 100% and specificity of 91.25% (NanoDrop spectrophotometer) to 98.75% (Qubit 3.0 single stranded assay or TaqMan real-time PCR).
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Cardiovascular Disease
- Normal
Platform:
Analyte Technology Platform DNA Fluorometry DNA Real-time qPCR DNA Spectrophotometry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Real-time qPCR Specific Technology platform NanoDrop spectrophotometer
Single-stranded Qubit 3.0 DNA assay
Double-stranded Qubit 3.0 DNA assay
TaqMan Copy Number Reference Assay TERT
Preaquisition Diagnosis/ patient condition ST elevated myocardial infarction
Healthy