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Impact of Preanalytical and Analytical Methods on Cell-Free DNA Diagnostics
Frontiers in Cell and Developmental Biology, 9 (2021); 686149. https://doi.org/10.3389/fcell.2021.686149
Krasić, Jure; Abramović, Irena; Vrtarić, Alen; Nikolac Gabaj, Nora; Kralik-Oguić, Saša; Katušić Bojanac, Ana; Ježek, Davor; Sinčić, Nino

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Krasić, J., Abramović, I., Vrtarić, A., Nikolac Gabaj, N., Kralik Oguić, S., Katušić Bojanac, A. ... Sinčić, N. (2021). Impact of Preanalytical and Analytical Methods on Cell-Free DNA Diagnostics. Frontiers in Cell and Developmental Biology, 9.. doi: 10.3389/fcell.2021.686149

Krasić, Jure, et al. "Impact of Preanalytical and Analytical Methods on Cell-Free DNA Diagnostics." Frontiers in Cell and Developmental Biology, vol. 9, 2021. https://doi.org/10.3389/fcell.2021.686149

Krasić, Jure, Irena Abramović, Alen Vrtarić, Nora Nikolac Gabaj, Saša Kralik Oguić, Ana Katušić Bojanac, Davor Ježek i Nino Sinčić. "Impact of Preanalytical and Analytical Methods on Cell-Free DNA Diagnostics." Frontiers in Cell and Developmental Biology 9 (2021). https://doi.org/10.3389/fcell.2021.686149

Krasić, J., et al. (2021) 'Impact of Preanalytical and Analytical Methods on Cell-Free DNA Diagnostics', Frontiers in Cell and Developmental Biology, 9. doi: 10.3389/fcell.2021.686149

Krasić J, Abramović I, Vrtarić A, Nikolac Gabaj N, Kralik Oguić S, Katušić Bojanac A, i sur.. Impact of Preanalytical and Analytical Methods on Cell-Free DNA Diagnostics. Frontiers in Cell and Developmental Biology [Internet]. 06.09.2021. [pristupljeno 15.03.2025.];9. doi: 10.3389/fcell.2021.686149

J. Krasić, et al., "Impact of Preanalytical and Analytical Methods on Cell-Free DNA Diagnostics", Frontiers in Cell and Developmental Biology, vol. 9, Rujan 2021. [Online]. Dostupno na: https://urn.nsk.hr/urn:nbn:hr:105:468697. [Citirano: 15.03.2025.]

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Podaci o radu
Naslov (engleski)Impact of Preanalytical and Analytical Methods on Cell-Free DNA Diagnostics
AutorJure Krasić
AutorIrena Abramović
AutorAlen Vrtarić
AutorNora Nikolac Gabaj
AutorSaša Kralik-Oguić
AutorAna Katušić Bojanac
AutorDavor Ježek
AutorNino Sinčić
Autorova ustanovaSveučilište u Zagrebu
Medicinski fakultet
(KATEDRA ZA HISTOLOGIJU I EMBRIOLOGIJU)
Autorova ustanovaSveučilište u Zagrebu
Medicinski fakultet
(KATEDRA ZA MEDICINSKU BIOLOGIJU)
Znanstveno / umjetničko
područje, polje i grana		BIOMEDICINA I ZDRAVSTVO
Temeljne medicinske znanosti
Citologija, histologija i embriologija
Sažetak (engleski)
While tissue biopsy has for the longest time been the gold-standard in biomedicine, precision/personalized medicine is making the shift toward liquid biopsies. Cell-free DNA (cfDNA) based genetic and epigenetic biomarkers reflect the molecular status of its tissue-of-origin allowing for early and non-invasive diagnostics of different pathologies. However, selection of preanalytical procedures (including cfDNA isolation) as well as analytical methods are known to impact the downstream results.... Više Calls for greater standardization are made continuously, yet comprehensive assessments of the impact on diagnostic parameters are lacking. This study aims to evaluate the preanalytic and analytic factors that influence cfDNA diagnostic parameters in blood and semen. Text mining analysis has been performed to assess cfDNA research trends, and identify studies on isolation methods, preanalytical and analytical impact. Seminal and blood plasma were tested as liquid biopsy sources. Traditional methods of cfDNA isolation, commercial kits (CKs), and an in-house developed protocol were tested, as well as the impact of dithiothreitol (DTT) on cfDNA isolation performance. Fluorimetry, qPCR, digital droplet PCR (ddPCR), and bioanalyzer were compared as cfDNA quantification methods. Fragment analysis was performed by qPCR and bioanalyzer while the downstream application (cfDNA methylation) was analyzed by pyrosequencing. In contrast to blood, semen as a liquid biopsy source has only recently begun to be reported as a liquid biopsy source, with almost half of all publications on it being review articles. Experimental data revealed that cfDNA isolation protocols give a wide range of cfDNA yields, both from blood and seminal plasma. The addition of DTT to CKs has improved yields in seminal plasma and had a neutral/negative impact in blood plasma. Capillary electrophoresis and fluorometry reported much higher yields than PCR methods. While cfDNA yield and integrity were highly impacted, cfDNA methylation was not affected by isolation methodology or DTT. In conclusion, NucleoSnap was recognized as the kit with the best overall performance. DTT improved CK yields in seminal plasma. The in-house developed protocol has shown near-kit isolation performance. ddPCR LINE-1 assay for absolute detection of minute amounts of cfDNA was established and allowed for quantification of samples inhibited in qPCR. cfDNA methylation was recognized as a stable biomarker unimpacted by cfDNA isolation method. Finally, semen was found to be an abundant source of cfDNA offering potential research opportunities and benefits for cfDNA based biomarkers development related to male reproductive health. Sakrij dio sažetka
Ključne riječi (engleski)
Jezikengleski
Vrsta publikacijeZnanstveni rad - Izvorni znanstveni rad
Status objaveObjavljen
Vrsta recenzijeRecenziran
Verzija publikacijeObjavljena verzija rada (izdavačev PDF)
Naslov časopisaFrontiers in Cell and Developmental Biology
Brojčani podacivol. 9, 686149
e-ISSN2296-634X
DOIhttps://doi.org/10.3389/fcell.2021.686149
URN:NBNurn:nbn:hr:105:468697
Datum objave publikacije2021-09-06
URL dokumentahttps://www.frontiersin.org/articles/10.3389/fcell.2021.686149/full
Vrsta resursaTekst
Prava pristupaOtvoreni pristup
Uvjeti korištenjaIkona licence Imenovanje 4.0 međunarodna (CC BY 4.0)
Datum i vrijeme pohrane2022-04-26 11:27:29
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