Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Rome, Italy.
PLoS One. 2013;8(1):e52391. doi: 10.1371/journal.pone.0052391. Epub 2013 Jan 14.
Epitheliotropic viruses can find their way into sewage. The aim of the present study was to investigate the occurrence, distribution, and genetic diversity of Human Papillomaviruses (HPVs) in urban wastewaters. Sewage samples were collected from treatment plants distributed throughout Italy. The DNA extracted from these samples was analyzed by PCR using five PV-specific sets of primers targeting the L1 (GP5/GP6, MY09/MY11, FAP59/64, SKF/SKR) and E1 regions (PM-A/PM-B), according to the protocols previously validated for the detection of mucosal and cutaneous HPV genotypes. PCR products underwent sequencing analysis and the sequences were aligned to reference genomes from the Papillomavirus Episteme database. Phylogenetic analysis was then performed to assess the genetic relationships among the different sequences and between the sequences of the samples and those of the prototype strains. A broad spectrum of sequences related to mucosal and cutaneous HPV types was detected in 81% of the sewage samples analyzed. Surprisingly, sequences related to the anogenital HPV6 and 11 were detected in 19% of the samples, and sequences related to the "high risk" oncogenic HPV16 were identified in two samples. Sequences related to HPV9, HPV20, HPV25, HPV76, HPV80, HPV104, HPV110, HPV111, HPV120 and HPV145 beta Papillomaviruses were detected in 76% of the samples. In addition, similarity searches and phylogenetic analysis of some sequences suggest that they could belong to putative new genotypes of the beta genus. In this study, for the first time, the presence of HPV viruses strongly related to human cancer is reported in sewage samples. Our data increases the knowledge of HPV genomic diversity and suggests that virological analysis of urban sewage can provide key information useful in supporting epidemiological studies.
嗜上皮性病毒可进入污水中。本研究旨在调查城市废水中人乳头瘤病毒（HPV）的发生、分布和遗传多样性。从分布在意大利各地的处理厂采集污水样本。从这些样本中提取的 DNA 使用五组针对 L1（GP5/GP6、MY09/MY11、FAP59/64、SKF/SKR）和 E1 区（PM-A/PM-B）的 PV 特异性引物通过 PCR 进行分析，根据先前针对黏膜和皮肤 HPV 基因型检测验证的方案。对 PCR 产物进行测序分析，并将序列与来自 Papillomavirus Episteme 数据库的参考基因组进行比对。然后进行系统发育分析，以评估不同序列之间以及样品序列与原型株之间的遗传关系。在所分析的 81%的污水样本中检测到与黏膜和皮肤 HPV 型相关的广泛序列。令人惊讶的是，在 19%的样本中检测到与肛门生殖器 HPV6 和 11 相关的序列，在两个样本中鉴定出与“高危”致癌 HPV16 相关的序列。在 76%的样本中检测到与 HPV9、HPV20、HPV25、HPV76、HPV80、HPV104、HPV110、HPV111、HPV120 和 HPV145 beta 乳头瘤病毒相关的序列。此外，一些序列的相似性搜索和系统发育分析表明，它们可能属于 beta 属的假定新基因型。在这项研究中，首次在污水样本中报告了与人癌症密切相关的 HPV 病毒的存在。我们的数据增加了 HPV 基因组多样性的知识，并表明对城市污水的病毒学分析可以提供支持流行病学研究的有用关键信息。
