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首段尿中生物标志物和 HPV DNA 的检测：收集量和 DNA 提取方法的影响。

Impact of Collection Volume and DNA Extraction Method on the Detection of Biomarkers and HPV DNA in First-Void Urine.

机构信息

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.

Novosanis nv, 2110 Wijnegem, Belgium.

出版信息

Molecules. 2021 Apr 1;26(7):1989. doi: 10.3390/molecules26071989.

DOI:10.3390/molecules26071989
PMID:33915837
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8036936/
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 ( < 0.05). Additionally, the potential of PhHV-1 as internal control to monitor FV urine collection, storage, and processing was demonstrated.

摘要

在过去的十年中，人们越来越认识到首段尿液（FV）作为一种非侵入性液体活检方法，用于检测人乳头瘤病毒（HPV）DNA 和其他生物标志物的潜力。在这项研究中，我们研究了初始尿液流的体积是否会影响生物标志物的分析性能。同时，我们评估了不同的 DNA 提取方案，并在尿液保存剂中引入了内部对照。25 名被诊断为高危 HPV 的女性使用三种不同体积（4、10、20 毫升）的 FV 尿液收集装置（Colli-Pee）收集了三个家庭采集的 FV 尿液样本。每个收集管中预先填充了含有疱疹病毒 1（PhHV-1）DNA 的尿液保存介质作为内部对照。比较了五种不同的 DNA 提取方案，然后进行 GAPDH 和 PhHV-1（qPCR）、HPV DNA 和 HBB（HPV 风险分析）以及 ACTB（甲基化特异性 qPCR）的 PCR。结果表明，收集量对人体和 HPV DNA 终点的影响有限。相比之下，不同 DNA 提取方法的人体终点的产量存在显著差异（<0.05）。此外，还证明了 PhHV-1 作为内部对照来监测 FV 尿液收集、储存和处理的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/669a/8036936/9fd7cd853660/molecules-26-01989-g004.jpg
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