江西一例iVDPV病例Ⅲ型脊髓灰质炎病毒基因特征分析  

Genetic Characterization of Type 3 Immunodeficiency Vaccine-derived Polioviruses in A Patient in Jiangxi Province,China

作　　者：郭琴 刘晓庆[3] 肖芳[3] 祝双利 王东艳 赵鹤鹤 路环环 肖金波 杨倩 李晓嫘 朱晖 冀天娇 王雯慧 何云 孙强 张勇 许文波[1,5] 严冬梅[1] GUO Qin;LIU Xiaoqing;XIAO Fang;ZHU Shuangli;WANG Dongyan;ZHAO Hehe;LU Huanhuan;XIAO Jinbo;YANG Qian;LI Xiaolei;ZHU Hui;JI Tianjiao;WANG Wenhui;HE Yun;SUN Qiang;ZHANG Yong;XU Wenbo;YAN Dongmei(National Polio Laboratory,WHO WPRO Regional Polio Reference Laboratory,National Health Commission Key Laboratory for Biosecurity,National Health Commission Key Laboratory for Medical Virology,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Dazhou Vocational College of Chinese Medicine,Dazhou 635000,China;Jiangri Center for Disease Control and Prevention,Nanchang 330029,China;Shandong First Medical University&Shandong Academy ofMedical Sciences,Jinan 271016,China;Center for Biosafety Mega-Science,Chinese Academy of Sciences,Beijing 102206,China)

机构地区：[1]国家脊髓灰质炎实验室、世界卫生组织西太平洋区脊髓灰质炎参比实验室、国家卫生健康委员会生物安全重点实验室、国家卫生健康委员会、医学病毒和病毒病重点实验室、中国疾病预防控制中心、病毒病预防控制所,北京102206 [2]达州中医药职业学院,达州635000 [3]江西省疾病预防控制中心,南昌330029 [4]山东第一医科大学,济南271016 [5]中国科学院生物安全大科学研究中心,北京102206

出　　处：《病毒学报》2023年第2期468-476,共9页Chinese Journal of Virology

基　　金：国家重点研发计划(项目号:2021YFF0703801),题目:微生物科学数据整合标准及共享服务平台建设;国家重点研发计划(项目号:2021YFC2302003),题目:病毒监测网络数据标准及数据平台建设;北京市自然科学基金(项目号:L192014),题目:EVA71疫苗大规模应用后对我国手足口病病原谱的影响及其保护效果评价研究。

摘　　要：分析江西一例免疫缺陷患者体内连续采集的15份粪便标本中分离到的Ⅲ型脊髓灰质炎病毒的全长VP1区基因特征。将从15份标本中分离到的14株Ⅲ型iVDPVs进行噬斑纯化,每个病毒随机挑取10个噬斑,接着进行逆转录‐聚合酶链反应扩增,测定获取到的154株iVDPVs全长VP1区序列。通过系统发育分析和BEAST程序探究iVDPV病毒的进化特征,估算其进化速率和口服脊灰减毒活疫苗(attenuated oral polio vaccine,OPV)初始感染时间。14株iVDPVs与Ⅲ型脊髓灰质炎减毒疫苗株(Sabin Ⅲ)核苷酸和氨基酸同源性分别为97.8%~98.7%和97.6%~98.3%。相较于Sabin Ⅲ,14株iVDPVs VP1区第54位氨基酸发生了A→V的突变,这可能导致温度敏感表型和衰减表型的改变。根据拓扑结构系统发育树被划分为3个Lineages,其中17049‐1‐8,17049‐2‐2和17049‐2‐9分别属于Lineage 1和Lineage 2,其余属于Lineage 3。具有Lineage 3序列特征的克隆是优势克隆,呈现出随时间持续分化的特征。BEAST程序估算的14株iVDPVs全长VP1区核苷酸平均进化速率为9.29×10‐3/位点/年(95%置信区间:2.81×10‐3‐1.59×10‐2),OPV初始感染时间为2012年11月18日。MCC树(maximum clade credibility tree)显示前4个遗传分支的分化仅用了40 d。14株Ⅲ型iVDPVs具有高度相关性,它们感染患儿后进化非常迅速,在慢性感染的早期便开始了谱系的分化。突变的不断积累和关键位点的改变可以引起iVDPVs神经毒力的变化,导致患儿出现AFP症状。iVDPV病例持续的排毒给脊灰根除目标的实现带来了巨大挑战,在脊髓灰质炎消灭的最后阶段及时调整免疫策略,继续维持较高的免疫覆盖率,保持iVDPV监测的灵敏性显得十分重要。To analyze the genetic characteristics of the complete VP1 regions of 14 type 3 immunodeficiency vaccine‐derived polioviruses (iVDPVs) isolated from 15 sequential stool specimens in a patient suffering from immunodeficiency in Jiangxi Province,China.Plaque purification was undertaken for 14 type 3 iVDPV isolates.Ten plaque‐purified viruses of each iVDPV isolate were selected randomly from each isolate for reverse transcription‐polymerase chain reaction (RT‐PCR) amplification and sequencing for the VP1 region.The full‐length VP1 region of 154 iVDPVs was obtained.Phylogenetic analysis and BEAST software were used to explore the evolutionary characteristics of iVDPVs and estimated the evolutionary rate and time to initial infection with a live attenuated oral polio vaccine (OPV).The homology of nucleotides and amino acids (AAs) between the 14 iVDPVs and a type 3 attenuated polio vaccine strain (Sabin 3) was 97.8%–98.7%and 97.6%–98.3%,respectively.Comparison of Sabin 3 and 14 iVDPVs showed an A→V mutation at AA54 in the VP1region,which is a key temperature‐sensitive and attenuation determinant.The phylogenetic tree was divided into three lineages based on topology:17049‐1‐8 was classified as“lineage 1”,17049‐2‐2 and 17049‐2‐9 were classified as“lineage 2”,and the remainder were classified as“lineage 3”.Clones with characteristics in lineage3 were dominant and showed a continuous evolutionary trend over time.The mean nucleotide evolution rate of the full‐length VP1 region of 14 iVDPVs estimated by BEAST was 9.29×10-3substitutions/site/year (95%HPD range:2.81×10-3to 1.59×10-2),and the initial OPV infection was on 18 November 2012.The maximum clade credibility (MCC) tree showed that the differentiation of the top‐four genetic branches took only40 days.The 14 type 3 iVDPVs were highly related,evolved rapidly after human infection and diverge into lineages at an early stage of chronic infection.Accumulation of mutations in key sites can increase the neurovirulence of iVDPVs

关 键 词：脊髓灰质炎病毒 免疫缺陷 口服脊灰减毒活疫苗 

分 类 号：R373.2[医药卫生—病原生物学]