苏州市一起聚集性人感染柯萨奇病毒A组4型基因特征  

Genetic characteristics of coxsackie A4 virus in a cluster of human infections in Suzhou

作　　者：董泽丰 徐智慧 王笛 吴学飞 雅雪蓉 夏瑜[1] 沈强[1] DONG Ze-feng;XU Zhi-hui;WANG Di;WU Xue-fei;YA Xue-rong;XIA Yu;SHEN Qiang(Laboratory of Pathogenic Microbiology,Suzhou Center for Disease Control and Prevention,Suzhou 215004,China)

机构地区：[1]苏州市疾病预防控制中心病原微生物检验科,苏州215004

出　　处：《中华疾病控制杂志》2023年第3期363-367,共5页Chinese Journal of Disease Control & Prevention

基　　金：苏州市重大疾病、传染病预防和控制关键技术(GWZX202202,GWZX202002);苏州市医学重点学科(SZXK202117);苏州市卫生青年骨干人才“全国导师制”培训项目(QNGG2022030);苏州市姑苏卫生人才计划培养项目(GSWS202099)。

摘　　要：目的 对苏州市一起人呼吸道感染不明病原的聚集性疫情病例咽拭子样本进行初步病原分子鉴定,为疫情处置提供技术支撑。方法 采集该起疫情病例咽拭子标本,实时荧光定量逆转录聚合酶链反应(Real-time reverse transcription polymerase chain reaction, Real-time RT-PCR)方法检测可引起聚集性呼吸道感染疫情的常见病原(甲/乙型流感病毒、副流感病毒、呼吸道合胞病毒、冠状病毒、鼻病毒、偏肺病毒和腺病毒)核酸,同时采用FilmArray全自动医用PCR分析系统进行多病原筛查。对病例咽拭子标本进行高通量测序,并用特异性引物探针检测柯萨奇病毒A组4型(coxsackievirus A4, CV-A4)进行验证。结果 该起疫情采样送检的18份咽拭子样本经Real-time RT-PCR方法检测,其中14份样本柯萨奇A4型病毒检测阳性。Real-time RT-PCR和高通量基因测序方法在病例咽拭子标本中均检出柯萨奇A4型病毒特异性基因片段。序列分析表明苏州该起疫情病毒株全基因组序列与NCBI数据库的MN964082病毒核酸一致性最高,为98.11%;VP1蛋白氨基酸序列与原型株AY421762一致性为98.03%,发现6个位点突变(T23V、A34T、S102A、A200T、I262V和Y285H)。结论 CV-A4可引起聚集性呼吸道感染疫情,对学龄儿童等免疫力较弱的人群应加强健康监测以便疫情早期处置。Objective This study aimed to conduct a preliminary molecular identification of pathogens in throat swab samples from a cluster of of cases with unexplained respiratory infections in Suzhou City. Methods Throat swab specimens of the outbreak cases were collected, and Real-time reverse transcription polymerase chain reaction(Real-time RT-PCR) method was employed to detect nucleic acid of common pathogens(influenza A/B viruses, parainfluenza viruses, respiratory syncytial viruses, coronaviruses, rhinoviruses, metapneumoviruses and adenoviruses) for clustered respiratory infections. Concurrently, FilmArray automatic medical PCR analysis system was used for multi-pathogen screening. High-throughput sequencing was performed on the throat swab specimens. And the sequenced results were verified by using specific kit to detect coxsackievirus A4(CV-A4). Results Among the 18 throat swab samples submitted for testing during this outbreak, 4 were positive for CV-A4 by Real-time RT-PCR. CV-A4 specific gene fragments were identified in the throat swab samples via both Real-time RT-PCR and high-throughput sequencing method. Sequence analysis indicated that the complete genome sequence of the virus strain involved in this outbreak in Suzhou City shared the highest similarity(98.11%) to the MN964082 viral nucleic acid in the NCBI database. The VP1 protein amino acid sequence demonstrated a 98.03% similarity to the prototype strain AY421762. Six mutation sites were identified(T23V, A34T, S102A, A200T, I262V, and Y285H) when compared to AY421762. Conclusions CV-A4 has the potential to cause clustered respiratory infections. Enhanced health monitoring is necessary for vulnerable groups, particularly school-aged children, to facilitate prompt detection and management of such outbreaks.

关 键 词：实时荧光定量逆转录聚合酶链反应 突变 基因测序 柯萨奇病毒A组4型 

分 类 号：R115[医药卫生—公共卫生与预防医学]