TY - JOUR
T1 - Impact of Collection Volume and DNA Extraction Method on the Detection of Biomarkers and HPV DNA in First-Void Urine
AU - Téblick, Laura
AU - Van Keer, Severien
AU - De Smet, Annemie
AU - Van Damme, Pierre
AU - Laeremans, Michelle
AU - Rios Cortes, Alejandra
AU - Beyers, Koen
AU - Vankerckhoven, Vanessa
AU - Matheeussen, Veerle
AU - Mandersloot, Renee
AU - Floore, Arno
AU - Meijer, Chris J L M
AU - Steenbergen, Renske D M
AU - Vorsters, Alex
N1 - Funding Information: Funding: This research was funded by H2020 Eurostars, grant number 12396. The funder did not play a role in this research. S. Van Keer is supported by a postdoctoral fellowship of the Research Foundation-Flanders (1240220N). Publisher Copyright: Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/4/1
Y1 - 2021/4/1
N2 - The potential of first-void (FV) urine as a non-invasive liquid biopsy for detection of human papillomavirus (HPV) DNA and other biomarkers has been increasingly recognized over the past decade. In this study, we investigated whether the volume of this initial urine stream has an impact on the analytical performance of biomarkers. In parallel, we evaluated different DNA extraction protocols and introduced an internal control in the urine preservative. Twenty-five women, diagnosed with high-risk HPV, provided three home-collected FV urine samples using three FV urine collection devices (Colli-Pee) with collector tubes that differ in volume (4, 10, 20 mL). Each collector tube was prefilled with Urine Conservation Medium spiked with phocine herpesvirus 1 (PhHV-1) DNA as internal control. Five different DNA extraction protocols were compared, followed by PCR for GAPDH and PhHV-1 (qPCR), HPV DNA, and HBB (HPV-Risk Assay), and ACTB (methylation-specific qPCR). Results showed limited effects of collection volume on human and HPV DNA endpoints. In contrast, significant variations in yield for human endpoints were observed for different DNA extraction methods (p < 0.05). Additionally, the potential of PhHV-1 as internal control to monitor FV urine collection, storage, and processing was demonstrated.
AB - The potential of first-void (FV) urine as a non-invasive liquid biopsy for detection of human papillomavirus (HPV) DNA and other biomarkers has been increasingly recognized over the past decade. In this study, we investigated whether the volume of this initial urine stream has an impact on the analytical performance of biomarkers. In parallel, we evaluated different DNA extraction protocols and introduced an internal control in the urine preservative. Twenty-five women, diagnosed with high-risk HPV, provided three home-collected FV urine samples using three FV urine collection devices (Colli-Pee) with collector tubes that differ in volume (4, 10, 20 mL). Each collector tube was prefilled with Urine Conservation Medium spiked with phocine herpesvirus 1 (PhHV-1) DNA as internal control. Five different DNA extraction protocols were compared, followed by PCR for GAPDH and PhHV-1 (qPCR), HPV DNA, and HBB (HPV-Risk Assay), and ACTB (methylation-specific qPCR). Results showed limited effects of collection volume on human and HPV DNA endpoints. In contrast, significant variations in yield for human endpoints were observed for different DNA extraction methods (p < 0.05). Additionally, the potential of PhHV-1 as internal control to monitor FV urine collection, storage, and processing was demonstrated.
KW - Cervical cancer
KW - Extractions
KW - First-void urine
KW - Genotyping
KW - HPV
KW - Human papillomavirus
KW - Optimization
KW - Self-sampling
UR - http://www.scopus.com/inward/record.url?scp=85105207740&partnerID=8YFLogxK
U2 - https://doi.org/10.3390/molecules26071989
DO - https://doi.org/10.3390/molecules26071989
M3 - Article
C2 - 33915837
SN - 1420-3049
VL - 26
JO - Molecules
JF - Molecules
IS - 7
M1 - 1989
ER -