为探讨上海市不同来源的柯萨奇病毒B组3型(coxsackievirus B3，CVB3)菌株的循环动态变化及基因特征，本研究对分离自上海市环境污水、健康儿童和急性弛缓性麻痹(acute flaccid paralysis，AFP)病例中的CVB3菌株进行VP1区基因序列测定，并对照全球 CVB3代表株进行序列相似性分析和系统发生学分析。结果显示，1989—2021年上海市不同来源的CVB3分离株共有120株，属于D、E基因亚型。上海CVB3分离株的核苷酸和氨基酸相似性差异较大，分别为78.7%～100%和93.3%～100.0%。2016年首次从上海环境污水中分离到E基因亚型CVB3，2021年再次分离到该基因亚型。2021年上海环境污水中的E基因亚型CVB3分离株与2020年的广东手足口病(hand，foot，and mouth disease，HFMD)病例CVB3分离株VP1 区的核苷酸相似性较高，但上海暂未有该基因亚型感染病例报道。因此，仍须长期对环境污水和不同肠道病毒感染病例进行监测，以提高肠道病毒监测的敏感性，为肠道病毒相关疾病的诊疗提供数据支撑。

To explore the dynamic changes and genetic characteristics of coxsackievirus B3 (CVB3) isolated from different sources in Shanghai, the full VP1 sequences of CVB3 isolates from environmental sewage surveillance, healthy children and acute flaccid paralysis (AFP) surveillance were amplified and sequenced in this study. Similarity and phylogenetic analysis of CVB3 isolates in Shanghai and global representative CVB3 strains were performed. The results showed that there were 120 CVB3 strains isolated from different sources in Shanghai from 1989 to 2021, belonging to D and E genotypes. The similarities of nucleotide and amino acid of Shanghai CVB3 isolates were from 78.7% to 100% and from 93.3% to 100.0%, respectively. In 2016, the genotype E CVB3 was firstly isolated from sewage in Shanghai, and it was isolated again in 2021. The nucleotides of genotype E CVB3 isolates from sewage in 2021 were similar to those of CVB3 isolates from hand, foot, and mouth disease (HFMD) cases in Guangdong in 2020. But, Shanghai has no reported infection cases of genotype E CVB3. It is still necessary to have a long-term surveillance for sewage and enterovirus cases to improve the sensitivity of enterovirus surveillance, and provide data support for the diagnosis and treatment of enterovirus-related diseases.