• Identification of Mucosal HPV in Wastewater from Tehran Hospitals
  • Romina Roshannia,1 Seyed Reza Mohebbi,2,* Seyed Masoud Hosseini,3 Shabnam Kazemian,4 Amir Sadeghi,5 Mohammad Reza Zali,6
    1. 1Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran2Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology
    2. Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
    3. Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran
    4. Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
    5. Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
    6. Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran


  • Introduction: Human papillomaviruses (HPVs) are a large and heterogeneous group of DNA viruses—with more than 200 identified to date. Viruses are typically classified as mucosal or cutaneous types. Mucosal subtypes, particularly HPV-16, HPV-18, HPV-31, HPV-53, HPV-56, and HPV-66, are recognized as high-risk because they have a proven association with oncogenesis, specifically cervical and some head and neck cancers. Furthermore, low-risk mucosal HPVs such as HPV-6, HPV-11, HPV-84, and HPV-89 occur frequently in benign lesions, i.e., anogenital warts. The majority of studies on mucosal HPVs so far have used clinical samples, but increased interest is focused on their possible occurrence in environmental matrices. Wastewater monitoring has proved to be an insightful tool for tracking viral spread among the population, and hospital effluents pose a threat as a specific problem in this sense—patients constantly secrete viral particles, and these effluents are a potential reservoir and point of disease-causing HPVs' transmission. Detection of mucosal HPV types in wastewater from hospitals is therefore important in environmental risk assessment and making efficient public health interventions. In this study, we explored the prevalence and distribution of mucosal HPV genotypes in wastewater from five Tehran hospitals with the goal of providing valuable information for future implementation of environmental surveillance of HPV.
  • Methods: We sampled fifty influent and effluent wastewater samples from five Tehran hospitals over a period of three months between December and February. All the samples were subjected to thorough filtration, and DNA was extracted with standard protocols adjusted to provide maximum integrity of nucleic acids for final analysis. Routine PCR was conducted using consensus MY/GP primers directed against a conserved segment within the HPV L1 gene. Negative controls were included in every PCR cycle to eliminate any chance of contamination. PCR-positive products were cleaned and mailed for Sanger sequencing. The resulting sequences were then cross-matched and compared with the NCBI HPV reference database for genotypic typing.
  • Results: 8 of 50 samples examined were positive for HPV DNA (16%). Three of them provided sequence of acceptable quality to permit direct genotyping. Mucosal types of HPV identified included HPV-16 and HPV-31—both oncogenic, high-risk types—and HPV-84—a low-risk type. Both clinically relevant high- and low-risk HPV genotypes were detected in hospital wastewater, highlighting the reservoir capacity for pathogenic viruses in hospital effluent. Detection of HPV-16, specifically, is significant considering that it is strongly linked with oropharyngeal and cervical carcinomas. These results show that routine wastewater treatment procedures will not completely remove viral genetic material, allowing residual HPV DNA to remain in treated effluent with potential environmental spread and inadvertent human exposure consequences.
  • Conclusion: Identification of mucosal HPV types within Tehran hospital wastewater demonstrates proof of the efficacy of wastewater surveillance as a public health monitoring tactic. The incidence of high-risk types—HPV-16 and HPV-31 in particular—is extremely high, which implies possible risk of insufficiently treated hospital discharge. On the other hand, wastewater-based epidemiologic methods can function as a useful early warning system, guiding water safety policy and infection control recommendations. In summary, this study emphasizes the necessity to complement clinical surveillance with environmental monitoring, particularly for pathogens of public health concern.
  • Keywords: Mucosal HPV, hospital wastewater, MY/GP primers, PCR, environmental monitoring, Tehran, wastewater-b