成都市锦江区污水中猴痘病毒的监测及基因特征分析

Monitoring and genetic analysis of monkeypox virus in sewage from Jinjiang District, Chengdu City

目的 调查成都市锦江区污水中猴痘病毒含量,并对其进行分型溯源。方法 2023年7~9月对成都市第九污水处理再生水厂未经消杀处理的污水进行24 h间断采样,共采集26个样本。用聚乙二醇沉淀法将水样从105 mL浓缩到0.6 mL后,提取核酸,用实时荧光聚合酶链反应(Polymerase Chain Reaction, PCR)法检测猴痘病毒,用猴痘病毒假病毒标准物质制备标准曲线计算相对浓度。对阳性的样本进行猴痘病毒全基因组扩增子测序。结果 8个样本检测出猴痘病毒阳性,检出率30.8%,均为西非分支Ⅱb C.1型。各分离株基因与报道的临床阳性株非常相似,与2022年猴痘疫情暴发的基因组密切相关。各分离株序列之间也非常相似,相似性为99%以上。结论 污水中的猴痘病毒都与报道的临床毒株高度相似,可以推定污水中的猴痘病毒均来自于患者或携带者的人体脱落物或排泄物。污水中猴痘病毒浓度与阳性报告病例的发生率基本呈正相关,表明污水监测是监测猴痘感染传播趋势的可行方法。

Objective Investigate the content of monkeypox virus in Sewage from Jinjiang District,Chengdu City,and conduct typing and tracing of its source.Methods From July to September 2023,24 hour sampling were conducted for the untreated Sewage of Chengdu No.9 wastewater Treatment and Reclaimed Water Plant,and collect a total of 26 samples.Concentrate the water sample from 105 mL to 0.6 mL using polyethylene glycol precipitation method,extract nucleic acid,detection of monkeypox virus using real-time fluorescence polymerase chain reaction(PCR)method,preparation of standard curves using monkeypox virus pseudovirus reference materials to calculate relative concentrations.Sequencing of the whole genome amplicon of monkeypox virus on positive samples.Results Eight samples tested positive for monkeypox virus,with a detection rate of 30.8%,all of which were West African branchⅡb C.1.The genes of each isolated strain were very similar to the reported clinically positive strains and closely related to the genome of the 2022 monkeypox outbreak.The sequences of each isolate were also very similar,with a similarity of over 99%.Conclusion The monkeypox virus in sewage is highly similar to the reported clinical strains,and it can be inferred that the monkeypox virus in sewage comes from the human excrement or feces of patients or carriers.The concentration of monkeypox virus in sewage is basically positively correlated with the incidence of positive reported cases,indicating that sewage monitoring is a feasible method for monitoring the transmission trend of monkeypox infection.