摘要
环境污水监测是掌握脊髓灰质炎(简称脊灰)病毒(Poliovirus, PV)在人群流行或携带状态的重要手段。为了解脊灰疫苗实施序贯免疫策略后山东省PV循环及遗传变异特征,我们于2019-2024年在山东省3个监测哨点开展环境污水PV监测,按月采集污水标本,浓缩后接种细胞进行病毒分离,提取分离物核酸后进行VP1完整编码区的RT-PCR扩增、Sanger测序和系统发生分析。共采集205份污水样本,收获208株PV,均为疫苗相关株,无PV2型病毒或疫苗衍生PV病毒(VDPV)。PV1型73株(占35.1%),与Sabin株相比VP1区核苷酸变异个数≤4,其中变异数为1个的毒株最多,占43.8%;PV3型135株(占64.9%),VP1区变异个数≤8,其中变异数为0个的毒株最多,占50.4%。共发现5株高变异株病毒,均为PV3型。PV1型VP1区nt-268位点A-T变异(35/73)和nt-270位点A-G变异(13/73)是变异最频繁的位点,两者联合导致65.8%(48/73)毒株在该位点发生I-M或I-L氨基酸变异。系统发生分析显示来自不同污水样本的变异数≥3的分离株没有聚集。本研究丰富了人群中存在的疫苗株PV的遗传变异特征,证实了序贯免疫策略实施后疫苗株PV仍存在发生变异和传播的风险,需进一步加强其监测和相关研究。
Environmental sewage surveillance is a critical tool for detecting poliovirus(PV)circulation or silent transmission within human populations.To investigate the circulation and genetic variation of PV following the implementation of the sequential immunization strategy in Shandong Province,China,environmental PV surveillance was conducted at three sentinel sites from 2019 to 2024.Sewage samples were collected monthly,concentrated,and subjected to virus isolation using cell culture.Nucleic acids were extracted from the isolates,and the complete VP1 coding region was amplified by RT-PCR,followed by Sanger sequencing and phylogenetic analysis.A total of 205 sewage samples were collected,yielding 208 PV isolates,all of which were vaccine-related strains.No type 2 PV or vaccine-derived polioviruses(VDPVs)were detected.Among the isolates,73 were type 1 PV(35.1%),all exhibiting≤4 nucleotide substitutions in the VP1 region compared to the Sabin strain,strains with a single mutation were most prevalent(43.8%).The remaining 135 isolates were type 3 PV(64.9%),with≤8 nucleotide substitutions in VPl;among these,strains without mutations were most common(50.4%).Five highly divergent strains-all type 3 PV-were identified.The most frequent mutations in the VP1 region of type 1 PV occurred at nt-268(A→T;35/73)and nt-270(A→G;13/73);the combination of these mutations resulted in amino acid substitutions from isoleucine(I)to methionine(M)or leucine(L)at this position in 65.8%(48/73)of isolates.Phylogenetic analysis revealed no clustering among isolates with≥3 nucleotide substitutions across different sewage samples.This study enhances understanding of the genetic variation of vaccine-related PV strains circulating in the population and confirms that,even after the adoption of the sequential immunization strategy,vaccine strains remain capable of genetic mutation and transmission.These findings underscore the need for continued surveillance and further research.
作者
王萌萌
纪峰
刘尧
林小娟
徐爱强
陶泽新
WANG Mengmeng;JI Feng;LIU Yao;LIN Xiaojuan;XU Aiqiang;TAO Zexin(School of Public Health,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan 250117,China;Shandong CenterforDiseaseControl andPrevention,Jinan250014,China)
出处
《病毒学报》
北大核心
2025年第3期733-740,共8页
Chinese Journal of Virology
基金
泰山学者青年专家计划项目(项目号:tsqn202103187),题目:基于同一健康理念和下一代测序技术新型病毒的发现、溯源和进化动态研究
山东省医药卫生科技重点项目(项目号:202412051234),题目:新型人类小RNA病毒的发现、遗传进化规律及其与人类疾病的关系。
关键词
脊灰病毒
环境监测
变异
系统进化
基因型
Poliovirus
Environmental surveillance
Genetic variation
Phylogenetic analysis
Genotype
