Effects of Pre-analytical Variables on Cell-free DNA Extraction for Liquid Biopsy
Laboratory Medicine Online
; : 45-56, 2019.
Article
ي En
| WPRIM
| ID: wpr-760491
المكتبة المسؤولة:
WPRO
ABSTRACT
BACKGROUND:
Extraction of cell-free DNA (cfDNA) is a key step for determining the quality of cfDNA-related molecular diagnostics. We evaluated the effect of sample containers and sample storage conditions on cfDNA extraction.METHODS:
The cfDNA extraction using the MagMAX Cell-Free DNA Isolation Kit from five healthy controls and five lung cancer patients was evaluated according to the type of sample container and storage conditions K2-EDTA container, <1, 6, 24, and 48 hr storage at 4℃ after immediate plasma separation; and Cell-Free DNA BCT container, <1, 3, 7, and 14 days stored at room temperature. Mutation analysis of EGFR exons 18–21 was performed. To assess the effect of a delay in centrifugation, EDTA whole blood samples from five healthy individuals were stored at 4℃ for 6, 12, and 24 hr before plasma separation.RESULTS:
There was no significant difference in the amount and nucleic acid size of cfDNA in both controls and patients with cancer when EDTA plasma was stored at 4℃ up to 48 hr. The amount and size of cfDNA in the BCT container were not different up to 7 days; however, the 14-day sample showed an increase in cfDNA concentration due to genomic DNA contamination. EGFR mutations were detected on EDTA containers up to 48 hr and with BCT containers up to 14 days. When EDTA whole blood was stored at 4℃ and plasma separation was delayed, the cfDNA concentration increased from 24 hr.CONCLUSIONS:
The cfDNA extraction was affected by the sample containers and storage conditions.Key words
النص الكامل:
1
الفهرس:
WPRIM
الموضوع الرئيسي:
Plasma
/
Biopsy
/
DNA
/
Centrifugation
/
Exons
/
Edetic Acid
/
Pathology, Molecular
/
DNA Contamination
/
Lung Neoplasms
نوع الدراسة:
Diagnostic_studies
المحددات:
Humans
اللغة:
En
مجلة:
Laboratory Medicine Online
السنة:
2019
نوع:
Article