Mucosal and cutaneous human papillomaviruses detected in raw sewages

PLoS One. 2013;8(1):e52391. doi: 10.1371/journal.pone.0052391. Epub 2013 Jan 14.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Humans
  • Italy
  • Molecular Sequence Data
  • Mucous Membrane / virology*
  • Papillomaviridae / genetics*
  • Papillomaviridae / isolation & purification*
  • Papillomavirus Infections / virology*
  • Phylogeny
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sewage / virology*
  • Skin / virology*

Substances

  • Sewage

Associated data

  • GENBANK/HE805665

Grants and funding

This work was partially funded by the Ministry of Health, AIDS projects 2009–2012, and partially supported by the joint Italia-USA project “Assessing the Impact of GIV Norovirus on Human Health: a Molecular Epidemiological Investigation on Environmental and Clinical Samples as a Basis for the Design of Novel Diagnostic Tools for an Emerging Pathogen”. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.