Comparison of methods for isolation and quantification of circulating cell-free DNA from patients with endometriosis

Hübner H, Lubrich H, Blum S, Antoniadis S, Lermann J, Ekici AB, Fasching P, Beckmann M, Rübner M, Burghaus S (2021)

Publication Type: Journal article

Publication year: 2021

Journal

Reproductive Biomedicine Online Elsevier

DOI: 10.1016/j.rbmo.2021.08.004

Abstract

Research question: Which is the optimal extraction method for isolating and quantifying circulating cell-free DNA (ccfDNA) from patients with endometriosis? Endometriosis is a common benign disease, associated with pain, infertility and reduced quality of life. Endometriosis is also a known risk factor for various cancers. Robust biomarkers for early detection and prediction of prognosis, however, are lacking. CcfDNA is an easy to obtain biomarker associated with prognosis of cancer patients and enables non-invasive analysis of somatic mutations. Recently, elevated levels of ccfDNA were detected in patients with endometriosis. Design: Two different ccfDNA extraction methods were compared: Maxwell RSC ccfDNA plasma kit (Maxwell) and QiAamp minElute ccfDNA mini kit (QIAamp). The ccfDNA and circulating mitochondrial DNA (mtDNA) quantities from 34 patients diagnosed with endometriosis were analysed. Fluorometric measurement and quantitative reverse transcription polymerase chain reaction (qRT-PCR) of short and long ALU and mtDNA fragments were used to quantiy ccfDNA. Results: The yield of ccfDNA isolated with the Maxwell method was significantly higher compared with the QIAamp method (P < 0.0001). Integrity of ccfDNA was significantly higher in the QIAamp isolate (P < 0.0001). Recovered mtDNA was not significantly different between both extraction methods used. Conclusions: The choice of extraction method can significantly influence the ccfDNA output and integrity. Both methods, however, enabled isolation of sufficient ccfDNA for further downstream applications. With this approach, isolation of ccfDNA could enable the non-invasive detection and analysis of somatic mutation within endometriosis tissue.

Authors with CRIS profile

Hanna Hübner Department of Obstetrics and Gynaecology Hannah Lubrich Lehrstuhl für Innere Medizin (Geriatrie) Simon Blum Department of Obstetrics and Gynaecology Sophia Antoniadis Department of Obstetrics and Gynaecology Arif Bülent Ekici Lehrstuhl für Humangenetik Peter Fasching Professur für Translationale Frauenheilkunde und Geburtshilfe Matthias Beckmann Lehrstuhl für Geburtshilfe und Frauenheilkunde Matthias Rübner Department of Obstetrics and Gynaecology Stefanie Burghaus Department of Obstetrics and Gynaecology

Involved external institutions

Klinikum Klagenfurt am Wörthersee AT Austria (AT)

How to cite

APA:

Hübner, H., Lubrich, H., Blum, S., Antoniadis, S., Lermann, J., Ekici, A.B.,... Burghaus, S. (2021). Comparison of methods for isolation and quantification of circulating cell-free DNA from patients with endometriosis. Reproductive Biomedicine Online. https://dx.doi.org/10.1016/j.rbmo.2021.08.004

MLA:

Hübner, Hanna, et al. "Comparison of methods for isolation and quantification of circulating cell-free DNA from patients with endometriosis." Reproductive Biomedicine Online (2021).

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