Department of Water Technology and Environmental Engineering, University of Chemistry and Technology Prague, Czechia.
Department of Water Technology and Environmental Engineering, University of Chemistry and Technology Prague, Czechia.
Sci Total Environ. 2023 Dec 1;902:166110. doi: 10.1016/j.scitotenv.2023.166110. Epub 2023 Aug 9.
Monkeypox virus (Mpxv) is a dsDNA virus that has become a global concern for human health in 2022. As both infected people and non-human hosts can shed the virus from their skin, faeces, urine and other body fluids, and the resulting sewage contains viral load representative of the whole population, it is highly promising to detect the spread of monkeypox virus in municipal wastewater. We established a methodology for sewage-based monitoring of Mpxv in Prague and analysed samples (n = 24) already early August-October of 2022 in a municipality with 1.4 million inhabitants that only reported 29 cumulative cases in this period. We isolated Mpxv DNA with the Wizard Enviro Total Nucleic Acid Kit, and thereafter detected Mpxv DNA using the EliGene® Monkeypox RT-PCR Kit. Prague wastewater was positive for Mpxv (in total 9 positive samples in periods with 1-9 new cases per week, coinciding with a weekly incidence of 0.07-0.64 per 100,000 inhabitants. The method for confirmation of wastewater positivity via semi-nested PCR and Sanger sequencing was successfully confirmed on positive controls including Mpxv particles and Mpxv-positive wastewater from the Netherlands. However, for Prague wastewater samples, amplification of Mpxv DNA via semi-semi-nested PCR was unsuccessful. This was probably due to extremely low case count, leading to the amplification of non-target bacterial DNA. Compared to other studies with much higher Mpxv prevalence, we show the outstanding sensitivity of our approach for monitoring the spread of monkeypox using wastewater.
猴痘病毒（Mpxv）是一种双链 DNA 病毒，在 2022 年已成为全球关注的人类健康问题。由于感染者和非人类宿主的皮肤、粪便、尿液和其他体液中都可能排出病毒，而由此产生的污水中含有代表整个人群的病毒载量，因此通过城市污水检测猴痘病毒的传播极具前景。我们在布拉格建立了一种基于污水的 Mpxv 监测方法，并于 2022 年 8 月初至 10 月分析了一个拥有 140 万居民的直辖市的样本（n=24），该直辖市在此期间仅报告了 29 例累计病例。我们使用 Wizard Enviro 总核酸试剂盒分离 Mpxv DNA，然后使用 EliGene®猴痘 RT-PCR 试剂盒检测 Mpxv DNA。布拉格污水中存在 Mpxv（总共 9 个阳性样本，每周有 1-9 个新病例，每周发病率为每 10 万人中 0.07-0.64 例。通过半巢式 PCR 和 Sanger 测序确认污水阳性的方法在包括 Mpxv 颗粒和荷兰 Mpxv 阳性污水在内的阳性对照中成功得到了确认。然而，对于布拉格污水样本，半巢式 PCR 扩增 Mpxv DNA 不成功。这可能是由于病例数极低，导致非目标细菌 DNA 的扩增。与其他 Mpxv 患病率高得多的研究相比，我们展示了使用污水监测猴痘传播的方法的出色灵敏度。
