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Article(id=1241522928743928650, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1241522919977841545, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202310262, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1697558400000, receivedDateStr=2023-10-18, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773931732593, onlineDateStr=2026-03-19, pubDate=1710000000000, pubDateStr=2024-03-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773931732593, onlineIssueDateStr=2026-03-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773931732593, creator=13701087609, updateTime=1773931732593, updator=13701087609, issue=Issue{id=1241522919977841545, tenantId=1146029695717560320, journalId=1227665162245664772, year='2024', volume='51', issue='5', pageStart='769', pageEnd='960', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773931730503, creator=13701087609, updateTime=1773931880386, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1241523548695622547, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1241522919977841545, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1241523548695622548, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1241522919977841545, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=916, endPage=920, ext={EN=ArticleExt(id=1241522929167553384, articleId=1241522928743928650, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Analysis of envelope protein gene characteristics of dengue virus in Hubei Province from 2014 to 2023, columnId=1228016572065837304, journalTitle=Modern Preventive Medicine, columnName=Experimental Technology and Applications, runingTitle=null, highlight=null, articleAbstract=
Objective

To analyze the characteristics of envelope protein (envelope, E) gene in dengue fever cases in Hubei Province from 2014 to 2023 and to trace the biological origin.

Methods

Serum samples of suspected dengue fever cases were collected, and nucleic acid detection and serum typing were carried out. Nucleic acid positive samples were amplified by RT-PCR and sequenced, and the gene sequences were obtained and analyzed by MEGA11.0 software.

Results

A total of 78 dengue nucleic acid positive cases were detected in Hubei Province from 2014 to 2023, and the E gene sequence of dengue virus (DENV) was successfully obtained in 44 samples, including 39 cases of serum type Ⅰ (DENV-Ⅰ) and 5 cases of serum typeⅡ (DENV-Ⅱ). Evolutionary analysis showed that 37 cases of DENV-Ⅰ were of gene type Ⅰ (G Ⅰ), distributed in three different branches, mainly close to the epidemic strains of Southeast Asian countries such as Guangzhou, Singapore, Indonesia, Thailand, and Myanmar, 2 cases were of gene type Ⅴ (GⅤ) were close to the epidemic strains of Guangzhou and India, and 5 cases of DENV-Ⅱ were of mixed type (Cosmopolitan type).

Conclusion

There was multitype dengue virus infection in Hubei Province from 2014 to 2023. The main epidemic strain was GⅠ subtype of DENV-Ⅰ, which was closely related to Guangzhou, China, and Southeast Asian countries. It is suggested that Hubei Province strengthen the prevention and control of cross-provincial and cross-border transmission of dengue fever and guard against the occurrence of severe dengue and death cases.

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目的

分析2014—2023年湖北省登革热感染病例包膜蛋白（envelope，E）基因特征并进行生物学溯源

方法

收集疑似登革热病例血清标本，对标本进行核酸检测及血清分型，核酸阳性标本采用RT-PCR扩增E基因并进行测序分型，获得基因序列用MEGA11.0软件进行基因特征分析。

结果

2014—2023年湖北省共检测登革热核酸阳性病例78份，成功获得44份标本的登革病毒（dengue virus, DENV）E基因序列，其中血清Ⅰ型(DENV-Ⅰ)39份，血清Ⅱ型(DENV-Ⅱ)5份；进化分析显示，DENV-Ⅰ中37例为基因Ⅰ型(GⅠ)，分布在三个不同分支上，主要与中国广州、新加坡、印度尼西亚、泰国和缅甸等东南亚国家流行株进化距离较近，2例为基因Ⅴ型(GⅤ)与中国广州和印度流行株进化距离较近；DENV-Ⅱ中5例均为混合型(cosmopolitan型)与输入地流行株进化距离较近。

结论

2014—2023年湖北省存在多型登革病毒感染，主要流行株为DENV-Ⅰ的GⅠ亚型，与中国广州和东南亚国家亲缘关系较近，提示湖北省应加强登革热跨省、跨境传播的防控，同时谨防登革重症及死亡病例的出现。

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邹文菁，E-mail:
, copyrightStatement=本刊刊出的所有文章不代表中华预防医学会和本刊编委会的观点，除非特别声明。, copyrightOwner=中华预防医学会和四川大学华西公共卫生学院, extLink=null, articleAbsUrl=null, sourceXml=BpJFuBEfRfu4dnuTr5ZvlQ==, magXml=7s0mHEB3FW8VvEYOO2y+wQ==, pdfUrl=null, pdf=tGSt92RY1dGV3Z4KAyQHUA==, pdfFileSize=1378003, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=SgHL9I0/fIi/7NXmtHHClw==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=MZkV+ySgL7n1T2SIcGUEFw==, mapNumber=null, authorCompany=null, fund=null, authors=

彭延（1976—），男，硕士，副主任技师，研究方向：病原微生物的检测及研究

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彭延（1976—），男，硕士，副主任技师，研究方向：病原微生物的检测及研究

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彭延（1976—），男，硕士，副主任技师，研究方向：病原微生物的检测及研究

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Notes:★for local cases; ▲for imported cases

, figureFileSmall=EfQz/keVbVfrPbpc5vcJ1Q==, figureFileBig=hWQWPniehsU15mlqQ9a+3w==, tableContent=null), ArticleFig(id=1241678307243118905, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522928743928650, language=CN, label=图1, caption=2014—2023年湖北省DENV-Ⅰ分离株E基因系统进化树

注：★为湖北省报告登革热本土病例；▲为湖北省报告登革热输入病例。

, figureFileSmall=EfQz/keVbVfrPbpc5vcJ1Q==, figureFileBig=hWQWPniehsU15mlqQ9a+3w==, tableContent=null), ArticleFig(id=1241678307805155656, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522928743928650, language=EN, label=Figure 2, caption=

Notes: ▲for imported cases

, figureFileSmall=Av+itgXg/wh3xipyG6ruUg==, figureFileBig=a2GTmBmjZnFM/5EnKQCkng==, tableContent=null), ArticleFig(id=1241678308581101907, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522928743928650, language=CN, label=图2, caption=2014—2023年湖北省DENV-Ⅱ分离株E基因系统进化树

注：▲为湖北省报告登革热输入病例。

, figureFileSmall=Av+itgXg/wh3xipyG6ruUg==, figureFileBig=a2GTmBmjZnFM/5EnKQCkng==, tableContent=null), ArticleFig(id=1241678310967660894, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522928743928650, language=EN, label=Table 1, caption=

Distribution of DENV serotypes in Hubei, 2014—2023

, figureFileSmall=null, figureFileBig=null, tableContent=
年份（年）标本数DENV核酸阳性DENV血清分型结果
阳性数阳性率（%）ⅠⅡⅢⅣ
201448612.56000
201517423.54000
201612541.73200
201712325.01200
201820735.04210
20191245040.348200
20236350.02100
合计2397832.668910
), ArticleFig(id=1241678311496143210, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522928743928650, language=CN, label=表1, caption=

湖北省2014—2023年DENV血清型分布

, figureFileSmall=null, figureFileBig=null, tableContent=
年份（年）标本数DENV核酸阳性DENV血清分型结果
阳性数阳性率（%）ⅠⅡⅢⅣ
201448612.56000
201517423.54000
201612541.73200
201712325.01200
201820735.04210
20191245040.348200
20236350.02100
合计2397832.668910
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湖北省2014—2023年登革病毒包膜蛋白基因特征分析
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彭延 , 蔡昆 , 李静 , 邹文菁
现代预防医学 | 实验技术及其应用 2024,51(5): 916-920
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现代预防医学 | 实验技术及其应用 2024, 51(5): 916-920
湖北省2014—2023年登革病毒包膜蛋白基因特征分析
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彭延, 蔡昆, 李静, 邹文菁
作者信息
  • 湖北省疾病预防控制中心，湖北 武汉 430079

    • 彭延（1976—），男，硕士，副主任技师，研究方向：病原微生物的检测及研究

通讯作者:

邹文菁，E-mail:
Analysis of envelope protein gene characteristics of dengue virus in Hubei Province from 2014 to 2023
Yan PENG, Kun CAI, Jing LI, Wen-jing ZOU
Affiliations
  • Hubei Center for Disease Control and Prevention, Wuhan, Hubei 430079, China
出版时间: 2024-03-10 doi: 10.20043/j.cnki.MPM.202310262
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摘要
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目的

分析2014—2023年湖北省登革热感染病例包膜蛋白（envelope，E）基因特征并进行生物学溯源

方法

收集疑似登革热病例血清标本，对标本进行核酸检测及血清分型，核酸阳性标本采用RT-PCR扩增E基因并进行测序分型，获得基因序列用MEGA11.0软件进行基因特征分析。

结果

2014—2023年湖北省共检测登革热核酸阳性病例78份，成功获得44份标本的登革病毒（dengue virus, DENV）E基因序列，其中血清Ⅰ型(DENV-Ⅰ)39份，血清Ⅱ型(DENV-Ⅱ)5份；进化分析显示，DENV-Ⅰ中37例为基因Ⅰ型(GⅠ)，分布在三个不同分支上，主要与中国广州、新加坡、印度尼西亚、泰国和缅甸等东南亚国家流行株进化距离较近，2例为基因Ⅴ型(GⅤ)与中国广州和印度流行株进化距离较近；DENV-Ⅱ中5例均为混合型(cosmopolitan型)与输入地流行株进化距离较近。

结论

2014—2023年湖北省存在多型登革病毒感染，主要流行株为DENV-Ⅰ的GⅠ亚型，与中国广州和东南亚国家亲缘关系较近，提示湖北省应加强登革热跨省、跨境传播的防控，同时谨防登革重症及死亡病例的出现。

登革病毒  /  包膜蛋白基因  /  序列分析
Objective

To analyze the characteristics of envelope protein (envelope, E) gene in dengue fever cases in Hubei Province from 2014 to 2023 and to trace the biological origin.

Methods

Serum samples of suspected dengue fever cases were collected, and nucleic acid detection and serum typing were carried out. Nucleic acid positive samples were amplified by RT-PCR and sequenced, and the gene sequences were obtained and analyzed by MEGA11.0 software.

Results

A total of 78 dengue nucleic acid positive cases were detected in Hubei Province from 2014 to 2023, and the E gene sequence of dengue virus (DENV) was successfully obtained in 44 samples, including 39 cases of serum type Ⅰ (DENV-Ⅰ) and 5 cases of serum typeⅡ (DENV-Ⅱ). Evolutionary analysis showed that 37 cases of DENV-Ⅰ were of gene type Ⅰ (G Ⅰ), distributed in three different branches, mainly close to the epidemic strains of Southeast Asian countries such as Guangzhou, Singapore, Indonesia, Thailand, and Myanmar, 2 cases were of gene type Ⅴ (GⅤ) were close to the epidemic strains of Guangzhou and India, and 5 cases of DENV-Ⅱ were of mixed type (Cosmopolitan type).

Conclusion

There was multitype dengue virus infection in Hubei Province from 2014 to 2023. The main epidemic strain was GⅠ subtype of DENV-Ⅰ, which was closely related to Guangzhou, China, and Southeast Asian countries. It is suggested that Hubei Province strengthen the prevention and control of cross-provincial and cross-border transmission of dengue fever and guard against the occurrence of severe dengue and death cases.

Dengue virus  /  Envelope protein gene  /  Sequence analysis
彭延, 蔡昆, 李静, 邹文菁. 湖北省2014—2023年登革病毒包膜蛋白基因特征分析. 现代预防医学, 2024 , 51 (5) : 916 -920 . DOI: 10.20043/j.cnki.MPM.202310262
Yan PENG, Kun CAI, Jing LI, Wen-jing ZOU. Analysis of envelope protein gene characteristics of dengue virus in Hubei Province from 2014 to 2023[J]. Modern Preventive Medicine, 2024 , 51 (5) : 916 -920 . DOI: 10.20043/j.cnki.MPM.202310262
正文
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登革热（dengue fever）是由登革病毒（dengue virus, DENV）引起在世界上传播范围最广的蚊媒传染病[1-2]。在过去50年间，发病率增加了30倍，并最终导致约2.5万人死亡[3-4]。目前登革热已成为全球范围内最严峻的公共卫生问题之一，同时造成极大经济负担。初步估算，我国在2019年登革热经济负担约为32亿元，占国内生产总值的0.04‰[5-7]。DENV属黄病毒科黄病毒属，基因组主要由三种结构蛋白（包膜、衣壳、膜）和7种非结构蛋白（NS1、NS2A、NS2B、NS3、NS4A、NS4B、、NS5）组成。包膜蛋白（envelope, E）是DENV的主要结构蛋白，影响病毒的进化过程及宿主间的传播[8]，因此常选择E基因进行病毒进化分析。湖北省夏季高温潮湿，人口密度大，对外交流日益增多，自2014年开始湖北省登革热输入病例逐年增多，在2019年发生了首起登革热本地感染疫情，登革热防控形势日趋严峻。但湖北省一直缺乏DENV纵向研究报告，本研究从基因水平分析湖北省2014—2023年DENV血清型及基因型特征，为湖北省登革热疫情防控提供科学依据。
1　材料与方法
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1.1　资料及标本来源
收集2014—2023年湖北省报告的疑似登革热病例急性期血清标本。病例资料来源于湖北省各市州疾控中心流行病学调查信息。
1.2　DENV检测和血清分型
用西安天隆科技有限公司病毒DNA/RNA提取试剂盒（磁珠法）提取总核酸，用上海伯杰医疗科技股份有限公司登革热通用及Ⅰ-Ⅳ型分型荧光定量检测试剂盒进行DENV核酸检测及血清型别鉴定，筛选核酸阳性标本。核酸提取和real-time RCR（RT-PCR）检测严格参照试剂说明书进行。
1.3　 E基因全长扩增及测序
将核酸阳性标本RNA用大连宝生物工程有限公司PrimeScript® One Step RT-PCR Kit Ver.2(Dye Plus)一步法试剂盒进行DENVE基因片段全长扩增，扩增引物及反应条件均参照文献[9]。引物合成、序列测定和拼接均委托武汉天一辉远生物科技有限公司完成。
1.4　序列分析
从美国国家生物技术信息中心（National Center for Biotechnology Information，NCBI）的基因库（GenBank）下载不同年代和地区流行的代表性毒株，涵盖DENV血清Ⅰ、Ⅱ型的所有基因亚型。其中DENV-Ⅰ参考株33株，DENV-Ⅱ参考株18株。用MEGA 11.0软件进行序列比对，并用邻接法构建E基因系统进化树，Bootstrap值取1 000。采用MegAlign软件进行核苷酸同源性分析。
2　结果
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2.1　基本情况
2014—2023年共收集登革热疑似病例血清标本239份，其中DENV核酸阳性78份，阳性率为32.6%，DENV分型RT-PCR检测鉴定为DENVⅠ-Ⅲ型，其中DNEVⅠ型68份，DENV-Ⅱ型9份，DENV-Ⅲ型1份。见表1。
2.2　确诊病例流行病学概况
确诊的78份病例输入性病例73例，本地感染6例。本地感染病例为一起暴发疫情。所有病例来源自全省15个地市州，其中武汉市确诊病例数最多为36例，其次为黄冈市12例，黄石7例，宜昌5例，随州和荆州各4例，十堰2例，剩余咸宁、襄阳、天门、仙桃、鄂州、恩施、荆门、孝感各1例。每年6—10月份为登革热发病高峰期，9月份病例数最多。78例确诊病例中男性56例（本地4例），女性22例（本地2例），男女比为2.55:1（本地病例男女比为2:1），年龄分布在20～67岁，年龄中位数为42岁。
2.3　DENV E基因进化分析
2.3.1　DENV E基因扩增与测序
78份DENV核酸阳性标本通过RT-PCR扩增E基因片段，测序拼接后，44份获得1 485bp的E基因全长序列，其中DENV-Ⅰ39份，DENV-Ⅱ5份。
2.3.2　DENV-ⅠE基因进化分析
DENV-Ⅰ型中37例为GⅠ亚型，2例为GⅤ亚型。进化分析显示37株GⅠ型核苷酸之间相似度在95.8%～100%之间，主要汇聚于3个大的分支上，分别设为A、B、C。A分支中除DG1-12-2019与2018年上海流行株以及DG1-19-2019和DG1-28-2019与2019年柬埔寨流行株进化距离较近外，其余24例2019年病例（包括6例本地疫情株）和1例2018年病例均与2019年广州流行株进化距离较近，相似度在99.21%～100%之间。B分支中1例DG1-31-2019与2020年留尼汪群岛病例进化距离较近，其余6例与中国广州、新加坡、印度尼西亚和泰国等东南亚国家流行株进化距离较近。相似度在99.47%～99.94%之间。分支C的3例与2018年缅甸和2019年广州流行株进化距离较近，相似度在99.80%～99.88%之间。2例GⅤ型集中在同一分支上，分别与2014年广州流行株和2011年印度流行株进化距离较近，相似度在99.93%～100%之间。见图1。
2.3.3　DENV-ⅡE基因进化分析
5例DENV-Ⅱ型病例基因分型均为cosmopolitan型,5例病例之间核酸序列相似度在99.83%～99.99%之间，其中DG2-2-2016和DG2-5-2018与新加坡流行株亲缘关系最近；其余3例分别与广州输入、印度尼西亚以及新喀里多尼亚流行株亲缘关系最近。见图2。
3　讨论
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湖北省属亚热带季风气候，夏季高温高湿持续时间长，利于蚊虫滋生，有研究显示湖北省3—11月均可监测到媒介伊蚊成蚊活动，给登革热的传播提供了充足的传播媒介[10-12]。加上近年气候不断变暖、城市化进程推进和劳务输出日益增多等诸多因素作用，使得登革热在湖北省流行呈现逐渐上升的趋势。2019年湖北省发生了首起登革热本地感染疫情[13-14]，提示湖北省已经具备适合DENV生存和循环传播的环境和媒介，而湖北省一直以来缺少对DENV血清、基因型及其特征的纵向研究。
本研究显示湖北省2014—2023年DENV存在3种血清型至少4种基因亚型,其中DENV-Ⅰ型一直保持持续流行的态势，为优势血清型，基因型以GⅠ亚型为主，这与近年中国登革热流行状况一致[15-16]。对比2014年至今湖北省登革热血清型还发现，湖北省DENV血清型存在由单一型向多种血清型并存的趋势，提示我们应该警惕DENV交叉传播的风险，预防重症及死亡病例的发生。同时有研究显示，2019年以后云南边境地区DENV主要流行血清型为DENV-3,DENV-1流行率相对降低，提示DENV-3正在取代以往优势DENV-1成为云南边境地区主要流行株[17]。边境地区血清型别的转换会否引起以后湖北省DENV主流血清型的转换有待于持续开展DENV病原学监测。
从构建的进化树可以看出湖北省DENV-Ⅰ的G1亚型分离株处于三个不同簇中，A分支中大部分的2019年样本聚集在一簇中，主要与中国广州2019年的参考毒株亲缘距离较近，提示这一簇样本可能来源于中国本土病例跨省输入。6例本土聚集性疫情病例也处于该分支中并与2019年广东流行株序列高度相似，提示湖北省本土疫情可能来源于广东省病例输入。分析2019年本土疫情前后湖北省登革热病例血清型及基因型发现，虽然2019年大部分病例和本土病例与2023年输入病例分处于A和B两个不同分支，但仍然属于DENV-Ⅰ的G1亚型，提示目前在湖北地区仍然以DENV-Ⅰ的G1亚型为优势血清型和基因亚型。C分支中少量2019年样本和1例2018年样本与中国广州和缅甸参考株聚集在一起，说明这一支样本可能来源于缅甸的输入并且引起中国本土流行。B分支中2014—2016年各1例，2019年2例和2023年1例处于同一分支，与印度尼西亚、新加坡、中国广州、泰国和法属留尼旺群岛亲缘关系较近，提示湖北省DENV1毒株输入来源复杂。值得一提的是DG1-31-Hubei-2019与2020年留尼旺群岛流行株亲缘关系接近，并与2014年广州流行株集中在同一分支，提示存在中国本土向外输出DENV的可能。本研究的2例基因ⅴ型与2008年，2011年印度和2014年广州聚集在同一分支，推测此毒株可能由印度输入到广州，并在广州形成本地及跨年传播进而输入湖北省。
2016—2019年的样本中有5例DENV-2病例完成基因测序。样本序列分析显示，虽然所有样本均为混合型（cosmopolitan型），但2014—2019年每年的DENV-2病例来源自不同国家。主要与新加坡、印度尼西亚等东南亚国家具有较近进化关系，包括同源性高的2017年广州病例也为马来西亚输入。再次提示近年DENV-2虽然处于低流行态势但湖北省DENV-2来源复杂，且受东南亚国家影响较大，今后应进一步开展DENV来源地研究。同时加强与东南亚国家接壤城市登革热监测，防止输入病例本地传播。
本研究存在一定不足。一是研究中各个年份样本数量存在一定不均衡性，同时由于新冠疫情影响，湖北省2020—2022年无登革热病例报告，实验结果可能存在一定偏差；二是研究中唯一1例DENV-3病例由于病毒载量低未成功获得序列。但本研究仍然从基因水平阐明了湖北省境内DENV的遗传进化特征，一定程度上反应了湖北省近年来DENV的流行及变化趋势，对制定相关防控策略提供指导。
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  • 国家自然科学基金区域创新发展联合基金重点项目(U21A20423)
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2024年第51卷第5期
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doi: 10.20043/j.cnki.MPM.202310262
  • 接收时间：2023-10-18
  • 首发时间：2026-03-19
  • 出版时间：2024-03-10
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  • 收稿日期：2023-10-18
基金
国家自然科学基金区域创新发展联合基金重点项目(U21A20423)
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    湖北省疾病预防控制中心，湖北 武汉 430079

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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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