Impact of Collection Volume and DNA Extraction Method on the Detection of Biomarkers and HPV DNA in First-Void Urine
Laura Téblick,
Severien Van Keer,
Annemie De Smet,
Pierre Van Damme,
Michelle Laeremans,
Alejandra Rios Cortes,
Koen Beyers,
Vanessa Vankerckhoven,
Veerle Matheeussen,
Renee Mandersloot,
Arno Floore,
Chris J. L. M. Meijer,
Renske D. M. Steenbergen,
Alex Vorsters
Affiliations
Laura Téblick
Centre for the Evaluation of Vaccination (CEV), Vaccine & Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, University of Antwerp, 2610 Wilrijk-Antwerp, Belgium
Severien Van Keer
Centre for the Evaluation of Vaccination (CEV), Vaccine & Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, University of Antwerp, 2610 Wilrijk-Antwerp, Belgium
Annemie De Smet
Centre for the Evaluation of Vaccination (CEV), Vaccine & Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, University of Antwerp, 2610 Wilrijk-Antwerp, Belgium
Pierre Van Damme
Centre for the Evaluation of Vaccination (CEV), Vaccine & Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, University of Antwerp, 2610 Wilrijk-Antwerp, Belgium
Michelle Laeremans
Novosanis nv, 2110 Wijnegem, Belgium
Alejandra Rios Cortes
Novosanis nv, 2110 Wijnegem, Belgium
Koen Beyers
Novosanis nv, 2110 Wijnegem, Belgium
Vanessa Vankerckhoven
Centre for the Evaluation of Vaccination (CEV), Vaccine & Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, University of Antwerp, 2610 Wilrijk-Antwerp, Belgium
Veerle Matheeussen
Department of Microbiology, Antwerp University Hospital (UZA), 2650 Edegem-Antwerp, Belgium
Renee Mandersloot
Self-Screen B.V, 1098 RX Amsterdam, The Netherlands
Arno Floore
Self-Screen B.V, 1098 RX Amsterdam, The Netherlands
Chris J. L. M. Meijer
Self-Screen B.V, 1098 RX Amsterdam, The Netherlands
Renske D. M. Steenbergen
Amsterdam UMC, Pathology, Cancer Center Amsterdam, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands
Alex Vorsters
Centre for the Evaluation of Vaccination (CEV), Vaccine & Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, University of Antwerp, 2610 Wilrijk-Antwerp, Belgium
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.