Meisal Roger, Rounge Trine Ballestad, Christiansen Irene Kraus, Eieland Alexander Kirkeby, Worren Merete Molton, Molden Tor Faksvaag, Kommedal Øyvind, Hovig Eivind, Leegaard Truls Michael, Ambur Ole Herman
Department of Microbiology and Infection Control, Akershus University Hospital, Lørenskog, Norway.
Department of Research, Cancer Registry of Norway, Oslo, Norway.
PLoS One. 2017 Jan 3;12(1):e0169074. doi: 10.1371/journal.pone.0169074. eCollection 2017.
Sensitive and specific genotyping of human papillomaviruses (HPVs) is important for population-based surveillance of carcinogenic HPV types and for monitoring vaccine effectiveness. Here we compare HPV genotyping by Next Generation Sequencing (NGS) to an established DNA hybridization method. In DNA isolated from urine, the overall analytical sensitivity of NGS was found to be 22% higher than that of hybridization. NGS was also found to be the most specific method and expanded the detection repertoire beyond the 37 types of the DNA hybridization assay. Furthermore, NGS provided an increased resolution by identifying genetic variants of individual HPV types. The same Modified General Primers (MGP)-amplicon was used in both methods. The NGS method is described in detail to facilitate implementation in the clinical microbiology laboratory and includes suggestions for new standards for detection and calling of types and variants with improved resolution.
人乳头瘤病毒（HPV）的灵敏且特异的基因分型对于基于人群的致癌性HPV类型监测以及疫苗效果监测至关重要。在此，我们将下一代测序（NGS）的HPV基因分型方法与一种既定的DNA杂交方法进行比较。在从尿液中分离出的DNA中，发现NGS的总体分析灵敏度比杂交法高22%。还发现NGS是最特异的方法，并且将检测范围扩展到了DNA杂交检测的37种类型之外。此外，NGS通过识别单个HPV类型的基因变异提供了更高的分辨率。两种方法都使用了相同的改良通用引物（MGP）扩增子。详细描述了NGS方法以促进其在临床微生物学实验室中的应用，包括关于提高分辨率的类型和变异检测及判定新标准的建议。
