Abstract
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.
Highlights
The potential of first-void (FV) urine as a suitable, non-invasive liquid biopsy for detection of human papillomavirus (HPV) DNA and other biomarkers to detect high-grade cervical abnormality, has already been reported [1,2,3,4,5,6,7,8]
We investigated whether the FV urine collection volume affects the detection of human and viral endpoints (i.e., glyceraldehyde 3-phosphate dehydrogenase (GAPDH), β-actin (ACTB), β-globin (HBB), HPV) by comparing three different FV
(i.e., internal control, collection volume, DNA extraction method) to investigate if we could improve the performance of FV urine as a liquid biopsy
Summary
The potential of first-void (FV) urine as a suitable, non-invasive liquid biopsy for detection of human papillomavirus (HPV) DNA and other biomarkers to detect high-grade cervical abnormality, has already been reported [1,2,3,4,5,6,7,8]. The rationale behind the use of FV urine is based on the fact that the initial stream of urine – first-void– washes away secretions that accumulate between the small labia and around the urethra opening, including mucus and debris from exfoliated cells from the vagina, cervix, and uterus [25]. This explains why the first fraction of a urine void contains significantly more HPV DNA than the subsequent part [12]. The currently used FV urine collection device (ColliPee® , Novosanis, Wijnegem, Belgium), prefilled with Urine Conservation Medium (UCM), collects the first fraction of urine in a total volume of 20 mL
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