Diverse fragment lengths dismiss size selection for serum cell-free DNA: a comparative study of serum and plasma samples.
Clin Chem Lab Med
; 58(9): 1451-1459, 2020 08 27.
Article
em En
| MEDLINE
| ID: mdl-32229658
ABSTRACT
Background:
The objective of this study was to determine the features of fragment length for circulating cell-free DNA (cfDNA) from plasma and serum samples.Methods:
Plasma and serum samples from different sources were randomly collected. Circulating cfDNA was extracted and purified by a precipitation-enriched and spin-column-based kit. The concentration of the purified DNA was immediately measured by a highly sensitive dsDNA quantitative assay, and then the fragment length was analyzed by capillary electrophoresis. The abundance of a specific fragment was estimated by the area under curve (AUC) for the fragment peak in the capillary electrophoresis.Results:
A total of 199 plasma and 117 serum samples were extracted and analyzed. The average yield of cfDNA from the serum samples (131.67 ng/mL) was significantly higher than that from the plasma samples (32.78 ng/mL, p < 0.001). The average abundance of the 20-400 bp fragments in plasma cfDNA (84.4%) was significantly higher than that of serum cfDNA (51.9%, p < 0.001). Fragment peaks in serum cfDNA always presented in regions around 190 bp, 430 bp, and 630 bp, but plasma cfDNA generally showed a sharp peak in the 165-190 bp region and a much lower peak in the 300Conclusions:
The fragment lengths of serum cfDNA and plasma cfDNA have very different features. Fragment size selection is suitable for plasma cfDNA but may not apply to serum cfDNA.Palavras-chave
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Base de dados:
MEDLINE
Assunto principal:
DNA Tumoral Circulante
Limite:
Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article