柯萨奇病毒A10型对利巴韦林耐药性的初步研究  

Preliminary Study on Resistance of Coxsackievirus A10 to Ribavirin

作　　者：孙甜甜 宗彦君 王蕊 李冀琛 刘莹 李晓亮 韩振志 于力恒 刘志军[1] 孙强[2,3] 张勇[2,3] SUN Tiantian;ZONG Yanjun;WANG Rui;LI Jichen;LIU Ying;LI Xiaoliang;HAN Zhenzhi;YU Liheng;LIU Zhijun;SUN Qiang;ZHANG Yong(Weifang Medical University,Weifang 261053,China;National Key Laboratory ofEpidemic Diseases Source Tracing,Early Warning and Intelligent Decision-making,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;National Polio Laboratory,National Health Commssion Key Laboratory for Medical Virology National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Shandong First Medical University,Jinan 250117,China;Virus Laboratory,Capital Institute of Pediatrics,Beijing Key Laboratory of Etiology of Childhood Viral Diseases,Beijing 100020,China)

机构地区：[1]潍坊医学院,潍坊261053 [2]传染病溯源预警和智能决策全国重点实验室,中国疾病预防控制中心病毒病预防控制所,北京102206 [3]国家脊髓灰质炎实验室,国家卫生健康委员会实验室生物安全重点实验室,国家卫生健康委员会医学病毒与病毒病重点实验室,中国疾病预防控制中心病毒病预防控制所,北京102206 [4]山东第一医科大学,济南250117 [5]首都儿科研究所病毒学研究室,儿童病毒病病原学北京市重点实验室,北京100020

出　　处：《病毒学报》2024年第2期337-345,共9页Chinese Journal of Virology

基　　金：国家重点研究发展计划项目(项目号:2021YFC2302003),题目:病毒监测网络数据标准及数据平台建设;山东省自然科学基金(项目号:ZR2019MC059),题目:巨细胞病毒基因UL138、IE1和IE2诱导的动脉粥样硬化机制研究。

摘　　要：近年来,全国范围内手足口病(Hand, foot, and mouth disease,HFMD)的感染率逐渐上升,其中柯萨奇病毒A组10型(Coxsackievirus A10,CV-A10)是主要病原体之一,利巴韦林是临床上治疗儿童手足口病的常用药物之一。为了评价利巴韦林在体外对CVA10的抑制效果,本研究中,我们随机选取了19条2009-2021年的不同地区和不同年代的有代表性的中国内地优势流行基因型C基因型的CVA10毒株,通过细胞毒性实验(Cell counting kit-8,CCK8)来检测利巴韦林在RD细胞上对CVA10的抑制率。将抑制率大于85%的CVA10定义为对利巴韦林敏感组,抑制率小于15%的CVA10定义为对利巴韦林耐药组。结果显示利巴韦林在0.78mmol/L浓度下可以对CVA10产生较好的抑制作用并且对细胞的毒性较低。19株CVA10中有4株表现出对利巴韦林耐药性,2株表现出对利巴韦林敏感性,剩余13株的敏感性在85%~15%之间。通过7日龄ICR乳鼠感染实验比较对利巴韦林敏感组和耐药组毒株的致病力差异,通过观察乳鼠临床评分、生存率、体重及组织取材的滴度测定结果均表明对利巴韦林敏感组CVA10在小鼠上的致病力比耐药组强。进一步使用MEGA 7.0软件对影响CVA10复制和翻译的2C区氨基酸序列进行差异性分析,以分析可能的耐药位点。结果显示对利巴韦林耐药组和敏感组CVA10的2C区的327位和328位氨基酸位点存在差异。本研究发现当前国内流行的CVA10存在广泛对利巴韦林耐药现象,其中部分CVA10对利巴韦林完全耐药。因此,有必要加强对CVA10耐药株的监测,以便更好地指导临床医生正确地选择抗病毒药物,为个体化的治疗提供理论依据。In recent years,the infection rate of hand,foot,and mouth disease(HFMD)has gradually increased nationwide,with Coxsackievirus Al0(CVA10)being one of the main pathogens.Ribavirin is one of the commonly used drugs for treatment of HFMD in children.In order to evaluate the clinical efficacy of ribavirin in the treatment of CVA10 infection,in this study,we randomly selected 19 representative CVA10 strains with dominant genotype C in mainland of China from 2009 to 2021 in different regions and different years,and detected the inhibition rate of ribavirin on CVA10 in RD cells through cell counting kit-8(CCK8).CVA10 strains with inhibition rate greater than 85%was defined as sensitive group to ribavirin,while CVA10 with inhibition rate less than 15%was defined as resistant group to ribavirin.The results showed that ribavirin had a good inhibitory effect on CVA10 at a concentration of o.78mmol/L and had low cytotoxicity to RD cells.Among the 19 CVA1O strains,four showed resistance to ribavirin,two showed sensitive to ribavirin,and the remaining thirteen showed sensitivity between 85% and 15%.The pathogenicity of the sensitive and resistant strains to ribavirin was compared in 7-day-old sucking mice infection experiments.The clinical scores,survival rate,body weight and viral titer measurements of tissue samples in suckling mice showed that the pathogenicity of the sensitive group CVA10 was stronger than that of the resistant group.Furthermore,the MEGA 7.0 software was used to perform differential analysis on the 2C region amino acid sequences that affect CVA10 replication and translation,in order to identify potential drug resistance sites.The results showed differences in the amino acid sites at positions 327 and 328 in the 2C region of CVA10 between the ribavirin-resistant group and the sensitive group.This study showed that the currently circulating CVA1O strains in China is widely resistance to ribavirin,with some CVAlo strains being completely resistant.Therefore,it is necessary to strengthen the monitoring of CVA10

关 键 词：柯萨奇病毒A10型 利巴韦林 耐药性 2C区 差异位点 

分 类 号：R373.2[医药卫生—病原生物学]