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Article(id=1241522776486507463, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1241522764012647140, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202305435, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1684944000000, receivedDateStr=2023-05-25, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773931696292, onlineDateStr=2026-03-19, pubDate=1704816000000, pubDateStr=2024-01-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773931696292, onlineIssueDateStr=2026-03-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773931696292, creator=13701087609, updateTime=1773931696292, updator=13701087609, issue=Issue{id=1241522764012647140, tenantId=1146029695717560320, journalId=1227665162245664772, year='2024', volume='51', issue='1', pageStart='1', pageEnd='192', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773931693318, creator=13701087609, updateTime=1773931808852, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1241523248643494379, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1241522764012647140, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1241523248643494380, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1241522764012647140, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=143, endPage=148, ext={EN=ArticleExt(id=1241522776973046779, articleId=1241522776486507463, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Analysis of molecular transmission network characteristics of newly confirmed HIV-1 infection in Jinan, columnId=1228016572065837304, journalTitle=Modern Preventive Medicine, columnName=Experimental Technology and Applications, runingTitle=null, highlight=null, articleAbstract=
Objective

To analyze the molecular network characteristics of the transmission relationship of newly confirmed human immunodeficiency virus type 1 (HIV-1) infection in Jinan, to provide basis for epidemic trend and prevention.

Methods

A total of 168 blood samples newly reported to be infected with HIV-1 in Jinan in 2021 were collected. The pol region gene of HIV-1 was amplified by nested PCR, the evolutionary tree was constructed by neighbor-joining method, and the molecular network was constructed by gene distance < 1.5%. The related factors of the network were analyzed. The mutation sites of drug resistance were analyzed by the online software tool of HIV drug resistance database of Stanford University.

Results

There were 9 subtypes in the sample sequence. The dominant subtypes were two epidemic recombination types circulating recombinant form (CRF), CRF01_AE subtype (41.7%, 70/168), and CRF07_BC subtype (36.3%, 61/168). The other subtypes were B, CRF68_01B,CRF55_01B, CRF65_CPX, CRF67_01B, CRF96_CPX, and C. When the gene distance was 1.50%, a total of 15 molecular clus ters were formed, with a total of 51 sequences, and the network entry rate was 30.4%. Surveillance drug-resistance mutations(SDRM) were monitored in 7 samples, with drug resistance transmission rate of 4.2%, focusing on drug resistance site mutations of nucleoside reverse transcriptase inhibitors and non-nucleoside reverse transcriptase inhibitors.

Conclusion

There are abundant HIV-1 subtypes in Jinan, mainly CRF01_AE and CRF07_BC subtypes. A molecular transmission network should be established for long-term monitoring to take targeted measures to block its spread.

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目的

分析济南市新确证人类免疫缺陷病毒1型（human immunodeficiency virus type 1, HIV-1）感染者传播关系的分子网络特征，为疫情流行趋势和防治提供依据。

方法

收集济南市2021年新报告的感染HIV-1的血液样本168份，通过巢式PCR扩增HIV-1的pol区基因，采用neighbor-joining方法构建进化树，基因距离<1.5%构建分子网络，对网络的相关因素进行分析。通过美国斯坦福大学HIV耐药数据库在线软件工具分析耐药突变位点。

结果

168份样本序列共9个亚型，优势亚型主要为两个流行重组型（circulating recombinant form, CRF），CRF01_AE亚型（41.7%，70/168）和CRF07_BC亚型（36.3%，61/168），其他亚型有B、CRF68_01B、CRF55_01B、CRF65_CPX、CRF67_01B、CRF96_CPX及C亚型。在基因距离为1.50%时，共形成了15个分子簇，入网序列共51条，入网率为30.4%。7份样本存在监测性耐药突变（surveillance drug-resistance mutations, SDRM),耐药传播率为4.2%，集中为核苷类反转录酶抑制剂和非核苷类反转录酶抑制剂耐药位点突变。

结论

济南市HIV-1亚型丰富，主要为CRF01_AE和CRF07_BC亚型。应构建分子传播网络对济南市进行长期监测，以采取针对性的措施阻断其传播。

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李新蕊，E-mail:；
刘岚铮，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=dXBoN8h2uWRPNV8tw76izA==, magXml=MjhzqskOJdXZUxn1suCkWw==, pdfUrl=null, pdf=3rrJb+TJmh1JN9l2VSvlUw==, pdfFileSize=722130, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=IyNLTiOrPOwZ4wacycoUTg==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=HXG6nGtsjXxXj+5/qf3r8Q==, mapNumber=null, authorCompany=null, fund=null, authors=

韩莹（1983—），女，硕士，副研究员，研究方向：病毒病

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韩莹（1983—），女，硕士，副研究员，研究方向：病毒病

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韩莹（1983—），女，硕士，副研究员，研究方向：病毒病

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Spatial and temporal transmission characteristics of two HIV strains CRF07 BC and CRF55 01B in China[D]. Beijing: Chinese center for disease control and prevention, 2021., articleTitle=Spatial and temporal transmission characteristics of two HIV strains CRF07 BC and CRF55 01B in China, refAbstract=null), Reference(id=1241677627065422578, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, doi=null, pmid=null, pmcid=null, year=2021, volume=100, issue=28, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[15], rfOrder=21, authorNames=Ma N, Chen XH, Zhao Y, journalName=Medicine (Baltimore), refType=null, unstructuredReference=Ma N, Chen XH, Zhao Y, et al. HIV-1 molecular transmission network among sexually transmitted populations in Liaoning Province, China[J]. Medicine (Baltimore), 2021, 100(28): e26640., articleTitle=HIV-1 molecular transmission network among sexually transmitted populations in Liaoning Province, China, refAbstract=null), Reference(id=1241677627191251705, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, doi=null, pmid=null, pmcid=null, year=2018, volume=24, issue=3, pageStart=241, pageEnd=245, 306, url=null, language=null, rfNumber=[16], rfOrder=22, authorNames=赵帅, 冯毅, 辛若雷, journalName=中国艾滋病性病, refType=null, unstructuredReference=赵帅，冯毅，辛若雷，等.应用分子传播网络研究北京男男性行为者HIV-1毒株的传播特征[J].中国艾滋病性病，2018，24（3）：241-245, 306., articleTitle=应用分子传播网络研究北京男男性行为者HIV-1毒株的传播特征, refAbstract=null), Reference(id=1241677627350635261, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, doi=null, pmid=null, pmcid=null, year=2018, volume=24, issue=3, pageStart=241, pageEnd=245, 306, url=null, language=null, rfNumber=[16], rfOrder=23, authorNames=Zhao S, Feng Y, Xin RL, journalName=Chinese Journal of AIDS & STD, refType=null, unstructuredReference=Zhao S, Feng Y, Xin RL, et al. Using molecular transmission network to explore the transmission characteristics of HIV-1 among men who have sex with men in Beijing[J]. Chinese Journal of AIDS & STD, 2018, 24(3): 241-245, 306., articleTitle=Using molecular transmission network to explore the transmission characteristics of HIV-1 among men who have sex with men in Beijing, refAbstract=null), Reference(id=1241677627497435909, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, doi=null, pmid=null, pmcid=null, year=2021, volume=10, issue=1, pageStart=1919, pageEnd=1930, url=null, language=null, rfNumber=[17], rfOrder=24, authorNames=Ge ZW, Feng Y, Zhang H, journalName=Emerging Microbes & Infections, refType=null, unstructuredReference=Ge ZW, Feng Y, Zhang H, et al.HIV-1 CRF07_BC transmission dynamics in China: two decades of National molecular surveillance[J]. Emerging Microbes & Infections, 2021, 10(1): 1919-1930., articleTitle=HIV-1 CRF07_BC transmission dynamics in China: two decades of National molecular surveillance, refAbstract=null), Reference(id=1241677627690373901, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, doi=null, pmid=null, pmcid=null, year=2020, volume=36, issue=9, pageStart=746, pageEnd=751, url=null, language=null, rfNumber=[18], rfOrder=25, authorNames=刘萌, 邢辉, 张玉琪, journalName=中国人兽共患病学报, refType=null, unstructuredReference=刘萌，邢辉，张玉琪，等.河北省治疗前人群HIV-1耐药毒株分子传播网络研究[J].中国人兽共患病学报，2020，36（9）：746-751., articleTitle=河北省治疗前人群HIV-1耐药毒株分子传播网络研究, refAbstract=null), Reference(id=1241677627770065683, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, doi=null, pmid=null, pmcid=null, year=2020, volume=36, issue=9, pageStart=746, pageEnd=751, url=null, language=null, rfNumber=[18], rfOrder=26, authorNames=Liu M, Xing H, Zhang YQ, journalName=Chinese Journal of Zoonoses, refType=null, unstructuredReference=Liu M, Xing H, Zhang YQ, et al. Molecular transmission of HIV-1 resistant strains among HIV-1 individuals before antiretroviral therapy in the Hebei Province[J]. Chinese Journal of Zoonoses, 2020, 36(9): 746-751., articleTitle=Molecular transmission of HIV-1 resistant strains among HIV-1 individuals before antiretroviral therapy in the Hebei Province, refAbstract=null), Reference(id=1241677627862340376, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, doi=null, pmid=null, pmcid=null, year=2017, volume=24, issue=2, pageStart=90, pageEnd=95, url=null, language=null, rfNumber=[19], rfOrder=27, authorNames=郝明强, 辛若雷, 王娟, journalName=国际病毒学杂志, refType=null, unstructuredReference=郝明强，辛若雷，王娟，等.北京市2016年新确证未治疗HIV感染者传播性耐药突变研究[J].国际病毒学杂志，2017，24（2）：90-95., articleTitle=北京市2016年新确证未治疗HIV感染者传播性耐药突变研究, refAbstract=null), Reference(id=1241677627963003677, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, doi=null, pmid=null, pmcid=null, year=2017, volume=24, issue=2, pageStart=90, pageEnd=95, url=null, language=null, rfNumber=[19], rfOrder=28, authorNames=Hao MQ, Xin RL, Wang J, journalName=International Journal of Virology, refType=null, unstructuredReference=Hao MQ, Xin RL, Wang J, et al. Surveillance of HIV transmitted drug resistance mutations in newly diagnosed treatment-naive patients in Beijing,2016[J]. International Journal of Virology, 2017, 24(2): 90-95., articleTitle=Surveillance of HIV transmitted drug resistance mutations in newly diagnosed treatment-naive patients in Beijing,2016, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1241677609826832394, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, xref=1., ext=[AuthorCompanyExt(id=1241677609835221003, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, companyId=1241677609826832394, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Jinan Center for Disease Control and Prevention, Jinan, Shandong 250021, China), AuthorCompanyExt(id=1241677609843609612, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, companyId=1241677609826832394, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.济南市疾病预防控制中心，山东 济南 250021)]), AuthorCompany(id=1241677609977827345, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, xref=2., ext=[AuthorCompanyExt(id=1241677609986215955, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, companyId=1241677609977827345, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.山东省疾病预防控制中心)])], figs=[ArticleFig(id=1241677619318542830, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=EN, label=Figure 1, caption=Phylogenetic tree of HIV-1, figureFileSmall=EukkUPM8ltcZ9htnsIMvqg==, figureFileBig=fUDRaPC2POxagzPqGmZBUA==, tableContent=null), ArticleFig(id=1241677619540840949, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=CN, label=图1, caption=HIV-1亚型系统进化树, figureFileSmall=EukkUPM8ltcZ9htnsIMvqg==, figureFileBig=fUDRaPC2POxagzPqGmZBUA==, tableContent=null), ArticleFig(id=1241677621042401791, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=EN, label=Figure 2, caption=Molecular transmission networks of HIV-1 subtype infected individuals in Jinan city at a genetic distance of 1.5%, figureFileSmall=tvZhmvHAZzuI3xYtB6oLcw==, figureFileBig=/RAq3uS1Svkxqm3rkO6PTQ==, tableContent=null), ArticleFig(id=1241677621214368264, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=CN, label=图2, caption=基因距离为1.5%时，济南市HIV-1亚型感染者的分子传播网络

注：CRF为流行重组型；图形大小表示病例与其他病例的相关程度。

, figureFileSmall=tvZhmvHAZzuI3xYtB6oLcw==, figureFileBig=/RAq3uS1Svkxqm3rkO6PTQ==, tableContent=null), ArticleFig(id=1241677621310837262, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=EN, label=Table 1, caption=

Information of 168 newly reported people with HIV-1

, figureFileSmall=null, figureFileBig=null, tableContent=
项目调查数（n=168）构成比（%）
性别
男16095.2
女84.8
年龄（岁）
≤359657.1
>357242.9
民族
汉16397.0
其他53.0
户籍
济南市15089.3
非济南市1810.8
婚姻状况
单身11467.9
已婚5432.1
文化程度
初中及以下5733.9
高中及以上11166.1
职业
商业服务4828.6
农民/工人4929.2
教师/学生137.7
家务或无业2917.3
其他2917.3
感染途径
异性性传播2917.3
同性性传播13982.7
CD4计数（个/μl）
<2003923.2
200～5008349.4
>5004627.4
亚型
CRF01_AE7242.9
CRF07_BC6035.7
其他3621.4
), ArticleFig(id=1241677621436666386, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=CN, label=表1, caption=

济南市168例新确证HIV-1型感染者的基本情况

, figureFileSmall=null, figureFileBig=null, tableContent=
项目调查数（n=168）构成比（%）
性别
男16095.2
女84.8
年龄（岁）
≤359657.1
>357242.9
民族
汉16397.0
其他53.0
户籍
济南市15089.3
非济南市1810.8
婚姻状况
单身11467.9
已婚5432.1
文化程度
初中及以下5733.9
高中及以上11166.1
职业
商业服务4828.6
农民/工人4929.2
教师/学生137.7
家务或无业2917.3
其他2917.3
感染途径
异性性传播2917.3
同性性传播13982.7
CD4计数（个/μl）
<2003923.2
200～5008349.4
>5004627.4
亚型
CRF01_AE7242.9
CRF07_BC6035.7
其他3621.4
), ArticleFig(id=1241677621533135388, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=EN, label=Table 2, caption=

Associated demographic factors of individuals clustering in Jinan HIV-1 molecular network ［n（%）］

, figureFileSmall=null, figureFileBig=null, tableContent=
项目调查例数（n=168）入网单因素分析OR值（95%CI）P值
性别
女8（4.8）2（25.0）1.00
男160（95.2）49（30.6）0.71（0.10～4.89）0.72
年龄（岁）
≤3596(57.1)29（30.2）1.00
>3572(42.9)22（30.6）0.98（0.51～1.91）0.96
民族
汉163（97.0）49（69.9）1.00
其他5（3.0）2（40.0）0.65(0.10～3.98)0.64
户籍
济南市150（89.3）48（32.0）1.00
非济南市18（10.8）3（16.7）2.35（0.65～8.52）0.19
婚姻状况
单身114（67.9）35（30.7）1.00
已婚54（32.1）16（29.6）0.95（0.47～1.93）0.89
文化程度
初中及以下57（33.9）18（31.6）1.00
高中及以上111（66.1）33（29.7）0.92（0.46～1.83）0.81
职业
家务或无业29（17.3）11（37.9）1.00
农民/工人49（29.2）11（22.4）0.47（0.17～1.30）0.15
商业服务48（28.6）15（31.3）0.74（0.28～1.96）0.55
教师/学生13（7.7）6（46.2）1.40（0.37～5.27）0.62
其他29（17.3）8（27.6）0.62（0.21～1.89）0.40
感染途径
异性性传播29（17.3）6（20.7）1.00
同性性传播139（82.7）45（32.4）2.04（0.65～6.40）0.22
CD4计数（个/μl）
<20039(23.2)11（28.2）1.00
200～50083(49.4)26（31.3）1.16（0.50～2.68）0.73
>50046(27.4)14（30.4）1.11（0.44～2.85）0.82
亚型
CRF07_BC6026（43.3）1.00
CRF01_AE7219（26.4）0.47（0.23～0.97）0.04
其他366（16.7）0.26（0.10～0.72）0.01
), ArticleFig(id=1241677621637992999, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=CN, label=表2, caption=

济南市HIV-1感染者入网率影响因素分析［n（%）］

, figureFileSmall=null, figureFileBig=null, tableContent=
项目调查例数（n=168）入网单因素分析OR值（95%CI）P值
性别
女8（4.8）2（25.0）1.00
男160（95.2）49（30.6）0.71（0.10～4.89）0.72
年龄（岁）
≤3596(57.1)29（30.2）1.00
>3572(42.9)22（30.6）0.98（0.51～1.91）0.96
民族
汉163（97.0）49（69.9）1.00
其他5（3.0）2（40.0）0.65(0.10～3.98)0.64
户籍
济南市150（89.3）48（32.0）1.00
非济南市18（10.8）3（16.7）2.35（0.65～8.52）0.19
婚姻状况
单身114（67.9）35（30.7）1.00
已婚54（32.1）16（29.6）0.95（0.47～1.93）0.89
文化程度
初中及以下57（33.9）18（31.6）1.00
高中及以上111（66.1）33（29.7）0.92（0.46～1.83）0.81
职业
家务或无业29（17.3）11（37.9）1.00
农民/工人49（29.2）11（22.4）0.47（0.17～1.30）0.15
商业服务48（28.6）15（31.3）0.74（0.28～1.96）0.55
教师/学生13（7.7）6（46.2）1.40（0.37～5.27）0.62
其他29（17.3）8（27.6）0.62（0.21～1.89）0.40
感染途径
异性性传播29（17.3）6（20.7）1.00
同性性传播139（82.7）45（32.4）2.04（0.65～6.40）0.22
CD4计数（个/μl）
<20039(23.2)11（28.2）1.00
200～50083(49.4)26（31.3）1.16（0.50～2.68）0.73
>50046(27.4)14（30.4）1.11（0.44～2.85）0.82
亚型
CRF07_BC6026（43.3）1.00
CRF01_AE7219（26.4）0.47（0.23～0.97）0.04
其他366（16.7）0.26（0.10～0.72）0.01
), ArticleFig(id=1241677621776405039, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=EN, label=Table 3, caption=

Distribution of association degrees of different subtypes in molecular propagation networks

, figureFileSmall=null, figureFileBig=null, tableContent=
亚型总数四分位间距中位数
CRF01_AE240.251
CRF07_BC1408.004
B80.51
), ArticleFig(id=1241677621902234169, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=CN, label=表3, caption=

分子传播网络内不同亚型关联程度的分布情况

, figureFileSmall=null, figureFileBig=null, tableContent=
亚型总数四分位间距中位数
CRF01_AE240.251
CRF07_BC1408.004
B80.51
), ArticleFig(id=1241677622049034813, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=EN, label=Table 4, caption=

Surveillance drug resistance mutations of newly reported people with HIV-1

, figureFileSmall=null, figureFileBig=null, tableContent=
样本名称NRTI SDRMs NNRTI SDRMsPI SDRMs
202109S0268—K101E—
202109S0470—K101E—
202109EW012D67N——
202109S0593D67E——
202109S0617—K103N—
202109S0632T69D——
202109S0647L74I, Y115F, M18K103N, V106M—
4V
), ArticleFig(id=1241677622170669636, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1241522776486507463, language=CN, label=表4, caption=

感染者存在的监测性耐药突变位点

, figureFileSmall=null, figureFileBig=null, tableContent=
样本名称NRTI SDRMs NNRTI SDRMsPI SDRMs
202109S0268—K101E—
202109S0470—K101E—
202109EW012D67N——
202109S0593D67E——
202109S0617—K103N—
202109S0632T69D——
202109S0647L74I, Y115F, M18K103N, V106M—
4V
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济南市新确证HIV-1型感染者的分子传播网络特征分析
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韩莹 1 , 潘晶 1 , 赵帅 2 , 滕婉莹 1 , 李欣 1 , 王美花 1 , 黄玉洁 1 , 韩秀云 1 , 李新蕊 1 , 刘岚铮 1
现代预防医学 | 实验技术及其应用 2024,51(1): 143-148
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现代预防医学 | 实验技术及其应用 2024, 51(1): 143-148
济南市新确证HIV-1型感染者的分子传播网络特征分析
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韩莹1, 潘晶1, 赵帅2, 滕婉莹1, 李欣1, 王美花1, 黄玉洁1, 韩秀云1, 李新蕊1 , 刘岚铮1
作者信息
  • 1.济南市疾病预防控制中心，山东 济南 250021
  • 2.山东省疾病预防控制中心

    • 韩莹（1983—），女，硕士，副研究员，研究方向：病毒病

通讯作者:

李新蕊，E-mail:；
刘岚铮，E-mail:
Analysis of molecular transmission network characteristics of newly confirmed HIV-1 infection in Jinan
Ying HAN1, Jing PAN1, Shuai ZHAO2, Wan-ying TENG1, Xin LI1, Mei-hua WANG1, Yu-jie HUANG1, Xiu-yun HAN1, Xin-rui LI1 , Lan-zheng LIU1
Affiliations
  • Jinan Center for Disease Control and Prevention, Jinan, Shandong 250021, China
出版时间: 2024-01-10 doi: 10.20043/j.cnki.MPM.202305435
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摘要
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目的

分析济南市新确证人类免疫缺陷病毒1型（human immunodeficiency virus type 1, HIV-1）感染者传播关系的分子网络特征，为疫情流行趋势和防治提供依据。

方法

收集济南市2021年新报告的感染HIV-1的血液样本168份，通过巢式PCR扩增HIV-1的pol区基因，采用neighbor-joining方法构建进化树，基因距离<1.5%构建分子网络，对网络的相关因素进行分析。通过美国斯坦福大学HIV耐药数据库在线软件工具分析耐药突变位点。

结果

168份样本序列共9个亚型，优势亚型主要为两个流行重组型（circulating recombinant form, CRF），CRF01_AE亚型（41.7%，70/168）和CRF07_BC亚型（36.3%，61/168），其他亚型有B、CRF68_01B、CRF55_01B、CRF65_CPX、CRF67_01B、CRF96_CPX及C亚型。在基因距离为1.50%时，共形成了15个分子簇，入网序列共51条，入网率为30.4%。7份样本存在监测性耐药突变（surveillance drug-resistance mutations, SDRM),耐药传播率为4.2%，集中为核苷类反转录酶抑制剂和非核苷类反转录酶抑制剂耐药位点突变。

结论

济南市HIV-1亚型丰富，主要为CRF01_AE和CRF07_BC亚型。应构建分子传播网络对济南市进行长期监测，以采取针对性的措施阻断其传播。

艾滋病  /  人类免疫缺陷病毒1型  /  分子传播网络  /  分子簇  /  耐药
Objective

To analyze the molecular network characteristics of the transmission relationship of newly confirmed human immunodeficiency virus type 1 (HIV-1) infection in Jinan, to provide basis for epidemic trend and prevention.

Methods

A total of 168 blood samples newly reported to be infected with HIV-1 in Jinan in 2021 were collected. The pol region gene of HIV-1 was amplified by nested PCR, the evolutionary tree was constructed by neighbor-joining method, and the molecular network was constructed by gene distance < 1.5%. The related factors of the network were analyzed. The mutation sites of drug resistance were analyzed by the online software tool of HIV drug resistance database of Stanford University.

Results

There were 9 subtypes in the sample sequence. The dominant subtypes were two epidemic recombination types circulating recombinant form (CRF), CRF01_AE subtype (41.7%, 70/168), and CRF07_BC subtype (36.3%, 61/168). The other subtypes were B, CRF68_01B,CRF55_01B, CRF65_CPX, CRF67_01B, CRF96_CPX, and C. When the gene distance was 1.50%, a total of 15 molecular clus ters were formed, with a total of 51 sequences, and the network entry rate was 30.4%. Surveillance drug-resistance mutations(SDRM) were monitored in 7 samples, with drug resistance transmission rate of 4.2%, focusing on drug resistance site mutations of nucleoside reverse transcriptase inhibitors and non-nucleoside reverse transcriptase inhibitors.

Conclusion

There are abundant HIV-1 subtypes in Jinan, mainly CRF01_AE and CRF07_BC subtypes. A molecular transmission network should be established for long-term monitoring to take targeted measures to block its spread.

AIDS  /  Human immunodeficiency virus type 1  /  Molecular transmission network  /  Molecular cluster  /  Drug resistance
韩莹, 潘晶, 赵帅, 滕婉莹, 李欣, 王美花, 黄玉洁, 韩秀云, 李新蕊, 刘岚铮. 济南市新确证HIV-1型感染者的分子传播网络特征分析. 现代预防医学, 2024 , 51 (1) : 143 -148 . DOI: 10.20043/j.cnki.MPM.202305435
Ying HAN, Jing PAN, Shuai ZHAO, Wan-ying TENG, Xin LI, Mei-hua WANG, Yu-jie HUANG, Xiu-yun HAN, Xin-rui LI, Lan-zheng LIU. Analysis of molecular transmission network characteristics of newly confirmed HIV-1 infection in Jinan[J]. Modern Preventive Medicine, 2024 , 51 (1) : 143 -148 . DOI: 10.20043/j.cnki.MPM.202305435
正文
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人类免疫缺陷病毒（human immunodeficiency virus, HIV）分子传播网络是近年来兴起的分子流行病学新方法，结合生物信息学及系统发育学来揭示HIV传播特征和流行规律，主要应用于HIV-1（HIV type 1）传播、耐药动力学和高危人群等方面的研究[1-4]。相比于同伴追踪、问卷调查、疾病监测等传统的流行病学手段，分子传播网络分析方法有其自身优点，将其应用于HIV-1传播关系的研究，可视化的传播网络图可以展现病毒的传播方向等规律，直观地展示HIV传播链的关键环节，便于我们有效的发现和控制真正的传染源[5]。为更好地了解济南市HIV-1的传播特征，本研究以济南市新确证HIV-1感染/AIDS患者为研究对象，结合人口学信息、序列、生物信息学及分子流行病学等方法，分析济南市新确证HIV-1感染者的人口学特点、流行重组亚型、入网率及耐药性，为济南市艾滋病疫情的防控提供充足的理论依据。
1　对象与方法
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1.1　研究对象
2021年济南市新报告的HIV感染者。纳入标准：首次确诊为HIV-1且未接受抗病毒治疗的感染者，年满18周岁，自愿参与调查研究，且签署知情同意书。最终168例HIV-1感染者参与调查，完成采血并成功测序。本研究通过济南市疾病预防控制中心伦理委员会审核批准（伦研批第LL20210906号），实施过程严格遵守相关的伦理学标准。
1.2　研究方法
1.2.1　病毒核酸提取及目标片段的扩增
采集感染者的全血样本，吸取部分全血样本24 h内完成CD4+T淋巴细胞计数。剩余全血样本1 500 r/min下离心15 min，分离血浆，-80℃冰箱保存备用。取200 μl血浆，按照QIAamp Viral RNA试剂盒说明书提取病毒RNA, QIAamp Viral RNA试剂盒来自德国Qiagen公司。参考文献[6]，采用反转录聚合酶链式反应（reverse transcription-polymerase chain reaction, RT-PCR）和巢式PCR对HIV-1病毒的pol区基因序列进行扩增，片段长度约1.2 kb。产物送北京诺赛基因组研究中心有限公司纯化并进行序列测定。
1.2.2　HIV-1基因亚型的判定
采用BioEdit7.1软件检查序列是否对齐，并进行手动校正。MEGAX软件对校正好的序列构建系统进化树并确定基因型别，进化树的构建采用neighbor-joining方法。
1.2.3　构建分子网络
比对好的序列导入HyPhy 2.2.4软件，采用Tamura-Nei93模型（TN93）计算序列两两之间的基因距离。根据《HIV传播网络监测和干预技术指南》的要求，按照1.5%基因距离阈值生成传播网络关系，利用Cytoscape 3.7.2软件计算相关网络参数并绘制网络图。
1.2.4　分析耐药性
将已经比对好的序列上传至美国斯坦福大学HIV耐药数据库中的在线软件（http://hivdb.stanford.edu/cpr/），进行监测性耐药突变（surveillancedrug resistance mutation, SDRM）分析，识别耐药突变位点。
1.2.5　统计学分析
研究对象信息录入Excel 2013软件，统计学分析采用IBM SPSS Statistics 21软件。用单因素logistic回归对入网率的影响因素进行分析，检验水准α=0.05。采用秩和检验分析不同亚型关联程度的差异。
2　结果
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2.1　基本特征及网络构成
168例患者中，男性160例（95.2%）；年龄以21~39岁为主，占58.3%；汉族163例（97.0%）；济南市户籍150例（89.3%）；未婚87例（51.8%）；教育程度大专及以上者74例（44.0%）；农民/工人49例（29.2%）；感染途径以同性性传播为主，占82.7%；CD4数量在200～500之间的83例（49.4%）。见表1。
168例样本共发现9个亚型，主要亚型为流行重组型01_AE亚型和CRF07_BC亚型，其它亚型包括B亚型22例（13.1%），CRF68_01B亚型3例，CRF55_01B亚型4例，CRF65_CPX亚型4例，CRF67_01B亚型1例，CRF96_CPX亚型1例，C亚型1例。见图1。
2.2　感染者入网率及其影响因素分析
基因距离为1.5%时，共形成15个分子簇，51条序列参与构成分子传播网络，入网率为30.4%。单因素分析与入网率相关的因素是亚型，亚型CRF07_BC的感染者入网率高于CRF01_AE亚型，显著高于其它亚型。CD4细胞数和感染途径与入网率无显著关联。见表2。
2.3　分子网络分析
图2入网序列包括三个亚型，CRF07_BC亚型、CRF01_AE亚型及B亚型，它们成簇的构成比分别为45.0%（27/60）、25.0%（18/72）和27.3%（6/22）。CRF07_BC在网络中包含5个传播簇，其中3个为男男同性传播感染，另两个传播簇中多数感染者为男男同性传播，少数为异性传播感染；两个传播簇包含的节点数大于5个，是网络中最大的两个传播簇，这两个传播簇中的病例均为济南本地病例；CRF01_AE在网络中包含8个传播簇，其中7个为男男同性性传播，一个既有男男同性传播又有异性传播感染；其节点数均小于5个，其传播簇中的人员均为男性；B在网络中包含2个传播簇，一个为男男同性性传播，一个既有男男同性传播又有异性传播感染；节点数均小于5。两两秩和检验表明，CRF07_BC的关联程度显著高于CRF01_AE（P<0.001），略高于B（P<0.05）,其余两组之间的关联程度没有显著差异。见表3。
2.4　耐药分析
168份样本中7份存在SDRM，耐药传播率为4.2%。其中单纯的NRTI耐药突变有3例，突变位点为D67N、D67E、T69D；单纯的NNRTI耐药突变有3例，其中两例的突变位点为K101E，一例突变位点为K103N；1例同时存在NRTI和NNRTI耐药突变，主要突变位点为L74I、Y115F、M184V、K103N、V106M；不存在蛋白酶抑制剂（protease inhibitor, PI）耐药突变，突变位点见表4。这7例耐药序列均不是入网序列，其中5例为济南本地病例，两例为外地病例，同时存在NRTI和NNRTI耐药的病例为安徽籍。
3　讨论
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第二次全国分子流行病学调查显示我国已成为HIV-1亚型最多的国家之一。我国已发现的有A、B、B'、C、D、E、F、G等10个亚型和多种不同的流行重组株[7]。本研究显示济南市HIV-1共存在7个亚型，主要亚型为CRF01_AE和CRF07_BC，与深圳、南京、宁夏等国内大部分地区研究结果保持一致[8-10]。CRF01_AE于上世纪70年代在中非共和国流行，80年代传入东南亚地区并造成广泛流行。1990年传入到我国云南省境内，通过不同流行路线，在国内不同地区蔓延扩散[11]。研究发现CRF01_AE的致病性比较强，且CRF01_AE感染者的病毒载量往往更高，使得该毒株在MSM人群中不仅传播速度更快，且患者的疾病进程也快于其他型别的感染者[12-13]。CRF07_BC于上世纪90年代起源于我国云南省，后通过性传播及静脉注射吸毒的方式在我国快速传播[14]，现已在各类人群中传播，成为主要优势毒株。
本研究中，当使用1.5%作为鉴定分子网络的基因距离阈值时，HIV-1分子传播网络的总入网率为30.4%。形成的分子网络，包含CRF01_AE、CRF07_BC和B三个亚型，且三个亚型独立成簇，不存在关联。说明三种亚型间的基因距离大于最适基因距离，他们相互之间的传播风险小于各亚型内部的传播风险，这与马宁在辽宁省的研究结果相吻合[15]。CRF07_BC的分子传播簇入网率高于其它两个亚型，在网络中的关联程度也高于其他两个亚型，传播关系更紧密、传播风险更高，这与赵帅在北京的研究结果一致，印证了CRF07_BC在中国北方男男性行为人群中广泛流行[16-17]。
济南市抗病毒治疗耐药突变率为4.2%，低于河北省、北京市等临近省市[18-19]。本研究发现济南市HIV-1感染者的潜在耐药突变位点主要集中为NRTI和NNRTI，未发现PI相关耐药位点。这可能与我国一线治疗药物以NRTI和NNRTI类为主，而PI类为二线药物，说明目前抗病毒治疗方案对于济南市新发感染者仍然适用。外地病例存在多重耐药，提示我们应加强耐药监测，防治外来病例引起的本地传播，出现多重耐药。
综上所述，济南市HIV-1基因型日趋复杂，存在多种亚型和重组型毒株，提醒我们要继续加强对济南市HIV感染者的分子流行病学研究，了解济南市新确证感染者的来源、分布及基因变化情况，掌握HIV-1的进化、变异规律。本研究收集到的样本有限，分析结论存在一定的局限性，后续还需进行进一步的扩展研究。
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2024年第51卷第1期
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doi: 10.20043/j.cnki.MPM.202305435
  • 接收时间：2023-05-25
  • 首发时间：2026-03-19
  • 出版时间：2024-01-10
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  • 收稿日期：2023-05-25
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    1.济南市疾病预防控制中心，山东 济南 250021
    2.山东省疾病预防控制中心

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2种不同金属材料的力学参数
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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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