Article(id=1321170497069495093, tenantId=1146029695717560320, journalId=1319373219989389464, issueId=1321170430187130949, articleNumber=null, orderNo=null, doi=10.11847/zgggws1143438, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1698595200000, receivedDateStr=2023-10-30, revisedDate=1733155200000, revisedDateStr=2024-12-03, acceptedDate=1737820800000, acceptedDateStr=2025-01-26, onlineDate=1792921192661, onlineDateStr=2026-10-25, pubDate=1739116800000, pubDateStr=2025-02-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792921192661, onlineIssueDateStr=2026-10-25, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792921192661, creator=13701087609, updateTime=1792921192661, updator=13701087609, issue=Issue{id=1321170430187130949, tenantId=1146029695717560320, journalId=1319373219989389464, year='2025', volume='41', issue='2', pageStart='129', pageEnd='256', issueExtLink='null', onlineDate='null', pubDate='1739116800000', pubDateStr='2025-02-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792921176716, creator='13701087609', updateTime=1792921176716, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=223, endPage=228, ext={EN=ArticleExt(id=1321170497287598902, articleId=1321170497069495093, tenantId=1146029695717560320, journalId=1319373219989389464, language=EN, title=Molecular epidemiologic characteristics and pretreatment drug resistance of HIV-1 in young and middle-aged first-diagnosed infections, 2019–2021 in Pudong New Area, Shanghai, columnId=1320427686514770171, journalTitle=Chinese Journal of Public Health, columnName=Survey and Report, runingTitle=null, highlight=null, articleAbstract=
Objective

To investigate the molecular epidemiological characteristics and drug resistance of HIV-1 among newly diagnosed antiretroviral therapy (ART)-naive infected individuals aged < 50 years in Pudong New Area, Shanghai, 2019–2021, and to provide evidence to refine local HIV prevention and control strategies.

Methods

Blood samples (n=374) were collected from individuals newly diagnosed with HIV infection between 2019 and 2021. Inclusion criteria were as follows: (1) confirmed first-time HIV-positive diagnosis by Western blot and without prior ART, (2) age less than 50 years, (3) voluntary participation, and (4) current residence or employment in Pudong New Area. Exclusion criteria were: (1) unsuccessful RNA extraction or sequencing, (2) mixed bases in the HIV-pol region sequence ≥5%, and (3) sequences shorter than 1 000 base pairs. The HIV-1 pol gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) for both pretreatment drug resistance genotyping and HIV-1 subtype determination, with subsequent analysis using the Stanford University HIV Drug Resistance Database for 302 successfully sequenced samples. Demographic and epidemiologic data were collected through structured in-person interviews.

Results

A total of 13 HIV-1 subtypes and unique recombinant forms (URFs) were identified among the 302 sequenced samples, of which 72.52% were from homosexual and 27.48% from heterosexual infections. The predominant strains were CRF07_BC (40.40%), CRF01_AE (36.09%) and CRF55_01B (8.28%). A significant variance in the distribution of viral subtypes was observed among individuals infected by different sexual transmission routes (χ2=22.161, P=0.003). However, the distribution of the prevalent strains CRF01_AE, CRF07_BC and CRF55_01B did not differ significantly between homosexual and heterosexual transmission (χ2=4.854, P=0.088), nor among the three age groups 15–24, 25–34 and 35–49 years (χ2=1.383, P=0.847). Resistance analysis showed a resistance rate of 5.96% in this population, with resistance levels ranging from low to intermediate to high. Non-nucleoside reverse transcriptase inhibitors (NNRTIs), nucleoside reverse transcriptase inhibitors (NRTIs) and protein inhibitors (PIs) accounted for 4.97%, 0.66% and 0.33% of resistance, respectively. The difference in resistance rates between subtypes was statistically significant (χ2=10.511, P=0.037). All cases (n=25) of the recombinant subtype CRF55_01BC had mutations at the resistance locus, resulting in an 88.00% incidence of mutations at potential resistance loci and a 12.00% incidence of resistance associated with moderate to high levels of resistance.

Conclusions

From 2019 to 2021, among the newly diagnosed HIV-1 infected individuals of young and middle-aged population in Pudong New Area, Shanghai, the genetic subtypes were predominantly CRF07_BC, CRF01_AE and CRF55_01B. However, new recombinant strains have emerged, indicating a trend toward diversification. In addition, varying degrees of pre-treatment drug resistance were observed in several subtypes.

, authors=Yan TANG1, 2, Jingjing LANG3, Qiuli XU1, 2, Huiqin FU1, 2, Panpan CHEN1, 2, Lipeng HAO1, 2, authorsList=Yan TANG, Jingjing LANG, Qiuli XU, Huiqin FU, Panpan CHEN, Lipeng HAO, authorCompany=null, correspAuthors=Lipeng HAO, authorNote=null, correspAuthorsNote=
HAO Lipeng, E-mail:
, copyrightStatement=Copyright © 2025 Chinese Journal of Public Health. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1321170497870607161, articleId=1321170497069495093, tenantId=1146029695717560320, journalId=1319373219989389464, language=CN, title=上海市浦东新区中青年感染人群HIV-1分子流行特征及耐药情况分析, columnId=1320427686770622717, journalTitle=中国公共卫生, columnName=调查报告与分析, runingTitle=null, highlight=null, articleAbstract=
目的

了解上海市浦东新区2019—2021年<50岁的中青年人群中未经抗病毒治疗的新确诊感染者/患者HIV-1分子流行病学特征及耐药情况,为艾滋病防治策略提供参考依据。

方法

收集2019—2021年浦东新区确诊实验室HIV-1抗体首次确诊阳性且人口学信息满足条件的347份<50岁人群血样本,采用反转录聚合酶链式反应(RT-PCR)扩增HIV-pol基因,进行治疗前耐药基因检测及 HIV-1亚型检测。

结果

获取302例HIV-pol区基因全序列片段,HIV-1流行簇主要有CRF07_BC(40.40%)、CRF01_AE(36.09%)、CRF55_01B(8.28%);感染途径为同性性传播(72.52%)和异性性传播(27.48%);不同性传播途径的病毒亚型分布差异有统计学意义(P=0.003)。HIV-1耐药率为5.96%,低中高度耐药均有出现,其中NNRTIs、NRTIs、PIs分别为4.97%、0.66%、0.33%;不同亚型之间的耐药率差异有统计学意义(P=0.037)。

结论

上海市浦东新区2019—2021年中青年人群新确诊HIV-1感染者,基因亚型主要以CRF07_BC、CRF01_AE 、CRF55_01B为主,但已出现新的重组毒株,呈现多元化发展趋势,且在多种亚型中出现不同程度的治疗前耐药。

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汤琰(1969—),副主任技师,本科,研究方向:微生物检测

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郝莉鹏,E-mail:
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figureFileSmall=cg3cgZg6xWUxzWwOCaCvTg==, figureFileBig=t1ZdTgMEP3bnuCt30OE5fQ==, tableContent=null), ArticleFig(id=1321170509337834355, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1321170497069495093, language=EN, label=Tab.1, caption=

Primers used for PCR sequencing and amplification of the pol gene in isolated HIV-1 strains with specific names, base compositions, orientations, positions, and lengths

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引物名称项目碱基构成及方向位置及长度/bp
4组扩增引物MAW-265′-TTGGAAATGTGGAAAGGAAGGAC-3′2027~2050
RT-215′-CTGTATTTCTGCTATTAAGTCTTTTGATGGG-3′3509~3539
PRO-15′-CAGAGCCAACAGCCCCACCA-3′2147~2166
RT205′-CTGCCAGTTCTAGCTCTGCTTC-3′3441~3462
5组测序引物PROS35′-GCCAACAGCCCCACCA-3′2151~2166
RTAS-qian5′-GGACCTACACCTGTCAAC-3′2484~2501
RTB5′-CCTAGTATAAACAATGAGACAC-3′2946~2968
PROC1S5′-GCTGGGTGTGGTATTCC-3′2826~2842
RT20S35′-GTTCTAGCTCTGCTTC-3′3441~3456
), ArticleFig(id=1321170509417526132, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1321170497069495093, language=CN, label=表1, caption=

HIV-1 PCR测序引物和扩增引物(pol基因)

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引物名称项目碱基构成及方向位置及长度/bp
4组扩增引物MAW-265′-TTGGAAATGTGGAAAGGAAGGAC-3′2027~2050
RT-215′-CTGTATTTCTGCTATTAAGTCTTTTGATGGG-3′3509~3539
PRO-15′-CAGAGCCAACAGCCCCACCA-3′2147~2166
RT205′-CTGCCAGTTCTAGCTCTGCTTC-3′3441~3462
5组测序引物PROS35′-GCCAACAGCCCCACCA-3′2151~2166
RTAS-qian5′-GGACCTACACCTGTCAAC-3′2484~2501
RTB5′-CCTAGTATAAACAATGAGACAC-3′2946~2968
PROC1S5′-GCTGGGTGTGGTATTCC-3′2826~2842
RT20S35′-GTTCTAGCTCTGCTTC-3′3441~3456
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Distribution of 302 newly diagnosed, treatment-naive HIV-1-infected individuals aged <50 years in Pudong New Area, Shanghai (2019–2021) by age, gender, marital status, educational attainment, household registration status, transmission route, and case identification method

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特征分类例数构成比/%特征分类例数构成比/%
年龄/岁15~243411.29受教育情况初中及以下8829.14
25~3414146.69高中或中专7324.17
35~4912742.05大专及以上14146.69
性别男性28293.38户籍情况上海市户籍24280.13
女性206.62非上海市户籍6019.87
婚姻状况未婚17557.95传播途径同性性传播21972.52
已婚有伴偶9631.79异性性传播8327.48
离异或丧偶3110.26发现途径主动检测发现12140.06
被动检测发现18159.93
), ArticleFig(id=1321170509585298294, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1321170497069495093, language=CN, label=表2, caption=

HIV/AIDS感染者/患者人口学及流行病学信息(N=302)

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特征分类例数构成比/%特征分类例数构成比/%
年龄/岁15~243411.29受教育情况初中及以下8829.14
25~3414146.69高中或中专7324.17
35~4912742.05大专及以上14146.69
性别男性28293.38户籍情况上海市户籍24280.13
女性206.62非上海市户籍6019.87
婚姻状况未婚17557.95传播途径同性性传播21972.52
已婚有伴偶9631.79异性性传播8327.48
离异或丧偶3110.26发现途径主动检测发现12140.06
被动检测发现18159.93
), ArticleFig(id=1321170509660795767, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1321170497069495093, language=EN, label=Tab.3, caption=

Distribution of HIV-1 gene subtypes among302 newly diagnosed, treatment-naive HIV-1-infected individuals aged < 50 years in Pudong New Area, Shanghai (2019–2021) by homosexual and heterosexual transmission routes

, figureFileSmall=null, figureFileBig=null, tableContent=
亚型同性性传播异性性传播各亚型占比
例数构成比/%例数构成比/%例数构成比/%
CRF07_BC8368.033931.9712240.40
CRF01_AE8779.822220.1810936.09
CRF55_01B1976.00624.00258.28
CRF08_BC116.67583.3361.99
CRF59_01B150.00150.0020.66
CRF67_01B375.00125.0041.32
CRF68_01B1100.0000.0010.33
CRF02_AGa00.0010.0010.33
A1 b00.0010.0010.33
B787.50112.5082.65
C00.003100.0030.99
URF1785.00315.00206.62
合计21983302
), ArticleFig(id=1321170509748876152, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1321170497069495093, language=CN, label=表3, caption=

302人/例中青年HIV感染者各基因亚型分布

, figureFileSmall=null, figureFileBig=null, tableContent=
亚型同性性传播异性性传播各亚型占比
例数构成比/%例数构成比/%例数构成比/%
CRF07_BC8368.033931.9712240.40
CRF01_AE8779.822220.1810936.09
CRF55_01B1976.00624.00258.28
CRF08_BC116.67583.3361.99
CRF59_01B150.00150.0020.66
CRF67_01B375.00125.0041.32
CRF68_01B1100.0000.0010.33
CRF02_AGa00.0010.0010.33
A1 b00.0010.0010.33
B787.50112.5082.65
C00.003100.0030.99
URF1785.00315.00206.62
合计21983302
), ArticleFig(id=1321170509845345145, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1321170497069495093, language=EN, label=Tab.4, caption=

Antiviral resistance profile, gene mutation site, number of strains, and potentially resistant drugsfor 51 strains isolated from 302 newly diagnosed, treatment-naive HIV-1-infected individuals aged < 50 years in Pudong New Area, Shanghai (2019–2021)

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耐药区间突变位点例数潜在耐药情况
PIsM46I1ATV/r、LPV/r
K20T1ATV/r
T74TP1ATV/r、DRV/r、LPV/r
F53FL1ATV/r
NRTIsK219KR2ABC、AZT、TDF
V75VI1ABC、AZT、FTC、3TC、TDF
A62V1ABC、AZT、FTC、3TC、TDF
NNRTIV106I2DOR、ETR、NVP、RPV
V179D10EFV、ETR、NVP、RPV
V179E31EFV、ETR、NVP、RPV
), ArticleFig(id=1321170509966979962, tenantId=1146029695717560320, journalId=1319373219989389464, articleId=1321170497069495093, language=CN, label=表4, caption=

具有潜在耐药的51例感染者/患者治疗前耐药突变位点

, figureFileSmall=null, figureFileBig=null, tableContent=
耐药区间突变位点例数潜在耐药情况
PIsM46I1ATV/r、LPV/r
K20T1ATV/r
T74TP1ATV/r、DRV/r、LPV/r
F53FL1ATV/r
NRTIsK219KR2ABC、AZT、TDF
V75VI1ABC、AZT、FTC、3TC、TDF
A62V1ABC、AZT、FTC、3TC、TDF
NNRTIV106I2DOR、ETR、NVP、RPV
V179D10EFV、ETR、NVP、RPV
V179E31EFV、ETR、NVP、RPV
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上海市浦东新区中青年感染人群HIV-1分子流行特征及耐药情况分析
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汤琰 1, 2 , 郎晶晶 3 , 许秋利 1, 2 , 傅惠琴 1, 2 , 陈盼盼 1, 2 , 郝莉鹏 1, 2, *, *
中国公共卫生 | 调查报告与分析 2025,41(2): 223-228
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中国公共卫生 |调查报告与分析 2025 , 41 (2) : 223 -228
上海市浦东新区中青年感染人群HIV-1分子流行特征及耐药情况分析
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汤琰1, 2, 郎晶晶3, 许秋利1, 2, 傅惠琴1, 2, 陈盼盼1, 2, 郝莉鹏1, 2, *, *
作者信息
  • 1上海市浦东新区疾病预防控制中心,上海 2001362
  • 2复旦大学浦东预防医学研究院,上海 200136
  • 3苏州大学附属儿童医院,江苏 215025
通讯作者:
郝莉鹏,E-mail:
作者简介:

汤琰(1969—),副主任技师,本科,研究方向:微生物检测

Molecular epidemiologic characteristics and pretreatment drug resistance of HIV-1 in young and middle-aged first-diagnosed infections, 2019–2021 in Pudong New Area, Shanghai
Yan TANG1, 2, Jingjing LANG3, Qiuli XU1, 2, Huiqin FU1, 2, Panpan CHEN1, 2, Lipeng HAO1, 2
Affiliations
  • 1Pudong District Center for Disease Control and Prevention, Shanghai 200136, China
  • 2Pudong Institute of Prevention Medicine, Fudan University, Shanghai 200136, China
  • 3Children's Hospital of Soochow University, Suzhou 215025, China
  • Corresponding Author:
    HAO Lipeng, E-mail:
出版时间: 2025-02-10 doi: 10.11847/zgggws1143438
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目的

了解上海市浦东新区2019—2021年<50岁的中青年人群中未经抗病毒治疗的新确诊感染者/患者HIV-1分子流行病学特征及耐药情况,为艾滋病防治策略提供参考依据。

方法

收集2019—2021年浦东新区确诊实验室HIV-1抗体首次确诊阳性且人口学信息满足条件的347份<50岁人群血样本,采用反转录聚合酶链式反应(RT-PCR)扩增HIV-pol基因,进行治疗前耐药基因检测及 HIV-1亚型检测。

结果

获取302例HIV-pol区基因全序列片段,HIV-1流行簇主要有CRF07_BC(40.40%)、CRF01_AE(36.09%)、CRF55_01B(8.28%);感染途径为同性性传播(72.52%)和异性性传播(27.48%);不同性传播途径的病毒亚型分布差异有统计学意义(P=0.003)。HIV-1耐药率为5.96%,低中高度耐药均有出现,其中NNRTIs、NRTIs、PIs分别为4.97%、0.66%、0.33%;不同亚型之间的耐药率差异有统计学意义(P=0.037)。

结论

上海市浦东新区2019—2021年中青年人群新确诊HIV-1感染者,基因亚型主要以CRF07_BC、CRF01_AE 、CRF55_01B为主,但已出现新的重组毒株,呈现多元化发展趋势,且在多种亚型中出现不同程度的治疗前耐药。

艾滋病病毒(HIV-1)  /  中青年人群  /  HIV-pol基因  /  基因亚型  /  上海市浦东区
Objective

To investigate the molecular epidemiological characteristics and drug resistance of HIV-1 among newly diagnosed antiretroviral therapy (ART)-naive infected individuals aged < 50 years in Pudong New Area, Shanghai, 2019–2021, and to provide evidence to refine local HIV prevention and control strategies.

Methods

Blood samples (n=374) were collected from individuals newly diagnosed with HIV infection between 2019 and 2021. Inclusion criteria were as follows: (1) confirmed first-time HIV-positive diagnosis by Western blot and without prior ART, (2) age less than 50 years, (3) voluntary participation, and (4) current residence or employment in Pudong New Area. Exclusion criteria were: (1) unsuccessful RNA extraction or sequencing, (2) mixed bases in the HIV-pol region sequence ≥5%, and (3) sequences shorter than 1 000 base pairs. The HIV-1 pol gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) for both pretreatment drug resistance genotyping and HIV-1 subtype determination, with subsequent analysis using the Stanford University HIV Drug Resistance Database for 302 successfully sequenced samples. Demographic and epidemiologic data were collected through structured in-person interviews.

Results

A total of 13 HIV-1 subtypes and unique recombinant forms (URFs) were identified among the 302 sequenced samples, of which 72.52% were from homosexual and 27.48% from heterosexual infections. The predominant strains were CRF07_BC (40.40%), CRF01_AE (36.09%) and CRF55_01B (8.28%). A significant variance in the distribution of viral subtypes was observed among individuals infected by different sexual transmission routes (χ2=22.161, P=0.003). However, the distribution of the prevalent strains CRF01_AE, CRF07_BC and CRF55_01B did not differ significantly between homosexual and heterosexual transmission (χ2=4.854, P=0.088), nor among the three age groups 15–24, 25–34 and 35–49 years (χ2=1.383, P=0.847). Resistance analysis showed a resistance rate of 5.96% in this population, with resistance levels ranging from low to intermediate to high. Non-nucleoside reverse transcriptase inhibitors (NNRTIs), nucleoside reverse transcriptase inhibitors (NRTIs) and protein inhibitors (PIs) accounted for 4.97%, 0.66% and 0.33% of resistance, respectively. The difference in resistance rates between subtypes was statistically significant (χ2=10.511, P=0.037). All cases (n=25) of the recombinant subtype CRF55_01BC had mutations at the resistance locus, resulting in an 88.00% incidence of mutations at potential resistance loci and a 12.00% incidence of resistance associated with moderate to high levels of resistance.

Conclusions

From 2019 to 2021, among the newly diagnosed HIV-1 infected individuals of young and middle-aged population in Pudong New Area, Shanghai, the genetic subtypes were predominantly CRF07_BC, CRF01_AE and CRF55_01B. However, new recombinant strains have emerged, indicating a trend toward diversification. In addition, varying degrees of pre-treatment drug resistance were observed in several subtypes.

HIV-1  /  young and middle-aged population  /  HIV-pol gene  /  genotype  /  Shanghai Pudong
汤琰, 郎晶晶, 许秋利, 傅惠琴, 陈盼盼, 郝莉鹏. 上海市浦东新区中青年感染人群HIV-1分子流行特征及耐药情况分析. 中国公共卫生, 2025 , 41 (2) : 223 -228 . DOI: 10.11847/zgggws1143438
Yan TANG, Jingjing LANG, Qiuli XU, Huiqin FU, Panpan CHEN, Lipeng HAO. Molecular epidemiologic characteristics and pretreatment drug resistance of HIV-1 in young and middle-aged first-diagnosed infections, 2019–2021 in Pudong New Area, Shanghai[J]. Chinese Journal of Public Health, 2025 , 41 (2) : 223 -228 . DOI: 10.11847/zgggws1143438
研究显示,男男性性行为人群(men who have sex with men, MSM)中HIV感染的比例较高,同时具有同性和异性性行为者占有一定的比例,已成为发达地区中青年人群感染HIV的主流群体[1–2]。尽管2015年开始推行诊断即治疗的艾滋病防治政策,但由于HIV感染后至抗病毒治疗的时间距离具有非常大的不确定性,如果晚期发现,传播给他人的概率较高,而且还有可能重组后使得亚型越来越复杂,同时部分感染者治疗后产生的耐药毒株直接传播给他人可导致新确诊的感染者在进行抗病毒治疗前就存在了耐药毒株(pretreatment drug resistance,PDR) [3–4]。当今由于网络的便捷和隐蔽性,通过网络寻找临时性伴侣已成为年轻人发生高危性行为的主要方式,从而突破了地域空间的局限性,与外区或外省形成了一个错综复杂的社会网络[5–6],同一区域内入网率相对中老年人群要低很多[7–9],给预防控制工作增加了很多难度。为了解上海市浦东新区中青年感染人群中的HIV-1分子流行病学特征和耐药情况,为浦东新区艾滋病防治策略提供参考依据,本研究收集了浦东新区2019—2021年<50岁中青年HIV感染者人群的血样本进行了分析。现将结果报告如下。
纳入标准(1)2019—2021年由浦东新区艾滋病中心实验室经蛋白印迹法(Western blot,WB)首次确诊HIV阳性的感染者/患者;(2)年龄<50岁;(3)自愿参与本研究并签署知情同意书;(4)现住址或户籍或工作地为浦东新区。排除标准:(1)RNA提取、扩增或测序失败;(2)HIV-pol区序列混合碱基≥5%;(3)序列长度<1 000 bp。最终成功测序302份。本研究经浦东新区疾病预防控制中心医学伦理委员会批准(批准号:202040477)。
自制调查问卷,有效问卷347份。由专业人员以面对面形式收集调查对象人口学及流行病学信息,包括年龄、居住地、户籍、学历、性取向、感染途径、发现途径等。
首次确诊后的调查对象全血5 mL,分离血浆后–70 ℃冰箱保存。采用LightCycler 480Ⅱ全自动核酸仪提取核糖核酸(RNA)(德国罗氏),按照操作说明书提取病毒RNA。采用套式反转录聚合酶链式反应(RT-PCR)和巢式PCR 扩增HIV-1 pol基因的蛋白酶编码区(RT),扩增试剂盒采用Prime Script TM One Step RNA RT-PCR Kit Ver.2和Takara Ex Taq。扩增片段大小约1.2 kb。扩增产物经电泳确认后送北京六合华大基因科技有限公司上海分公司进行扩增产物的纯化和测序,测序引物和扩增引物见表1。
在线工具HIV-BLAST( https://www.hiv.lanl.gov/content/sequence/BASIC_BLAST/basic_blast.Html) 进行初步分型。同时从HIV sequence database 网站下载HIV-1不同亚型流行毒株的参考序列,使用BioEdit v7.0.5.3 对pol 区片段序列和参考序列进行手动调整和比对,再利用FastTreev1.4.4中最大似然(maximum likelihood,ML)法构建系统进化树并根据其聚类结果判定中青年人群HIV-1基因亚型分布特征。流行簇定义:(1)各传播簇中的节点值(SH test ratio)≥0.8;(2)所含同一亚型基因序列数量>20条。流行簇中的分支定义:(1)各分支中的节点值(bootstrap≥0.9[6] ; (2)各分支内平均基因距离≤0.030 [7]。
运用美国斯坦福大学HIV-1耐药数据库进行治疗前耐药(PDR)分析。根据数据库系统评分将耐药程度分为:敏感(S)、潜在耐药(P)、低度耐药(L)、中度耐药(I)和高度耐药(H)5个水平,其中 L、I 和 H水平判定毒株为耐药毒株。
早期发现:根据2个不同时间点(天)血浆样本的确证条带数增加;或首次确证为阴性/不确定后随访抗体确证阳性,同时结合流行病学信息;或确证条带显示早期特征、同时结合流行病学特征及HIV检测史;晚期发现:根据2个不同时间点(天)血浆样本的确证条带衰减或首次确证为不确定后随访核酸确证阳性,同时结合流行病学及CD4+T基数<200 cell/μL;或确证条带显示晚期特征同时结合流行病学及CD4+T基数<200 cell/μL。主动发现:高危行为后主动前往医疗机构咨询检测发现;被动发现是因其他疾病就医后由医生评估检测或性伴侣阳性后由医生通知检测或术前产前筛查及其它监测中发现。口交传播的鉴定:HIV感染前只发生过口交行为,或HIV检测阴性后的最近一段时间内无肛交而只有口交行为,流调显示急性期以及确证条带显示早期特征。如果同一时间段内既有口交又有肛交行为,感染途径默认为通过肛交传播。
采用SPSS 16.0软件进行统计学分析。使用率或构成比描述,采用χ2检验或Fisher精确概率法进行统计学检验。检验水准α=0.05。
共收集调查问卷及对应的血样本347份,最终成功测序302例HIV/AIDS新确证感染者/患者。
302例HIV/AIDS感染者/患者传播途径包括同性性传播和异性性传播,而家庭配偶间传播占异性性传播的8.43%(7/83);早期和晚期发现分别占比31.12%(94例)、17.22%(52例)。高危性行为期间具有跨省、跨区临时性伴侣的分别占比24.17%(73例)、40.39%(122例)。同性性传播人群同一时期内既有同性性行为又有异性性行为者占比17.35%(38/219),具有固定同性性伴侣者占比4.57%(10/219),既有固定又有临时同性性伴侣者占比7.31%(16/219),通过口交传播者占比3.20%(7/219)。
302份HIV/AIDS的HIV-pol基因序列,确定共存在13种亚型及独特重组型(unique recombinant forms,URFs),其中CRF01_AE(36.09%,109/302)、CRF07_BC(40.40%,122/302)、CRF55_01B(8.28%,25/302)为浦东地区此人群的主要流行毒株。不同性传播途径的病毒亚型总体分布差异具有统计学意义(χ2=22.161,P=0.003),但流行毒株CRF01_AE、CRF07_BC、CRF55_01B分别在同性与异性性传播之间的分布特征差异无统计学意义(χ2=4.854,P=0.088)。
除CRF08_BC及C亚型以异性性传播为主外,其余亚型均以同性性传播为主。流行毒株CRF07_BC、CRF01_AE、CRF55_01B 同性性传播人群同时具有异性性行为者分别占比20.48%(17/83)、13.79%(12/87)、15.79%(3/19),三者差异无统计学意义(χ2=1.424,P=0.491);三者在15~24、25~34、35~49岁组占比分别为13.12%(16/122)、49.18%(60/122)、37.70%(46/122);11.01%(12/109)、45.87%(50/109)、43.12%(47/109);16.00%(4/25)、40.00%(10/25)、44.00%(11/25),其在3个年龄段之间的分布特征差异无统计学意义(χ2=1.383,P=0.847),同时其分布特征与3个年龄段人数分布特征之间差异均无统计学意义(均P>0.05),由此表明流行毒株CRF07_BC、CRF01_AE和CRF55_01B在3个年龄段之间分布比较均衡。
进化树分析显示,各株均按照基因型别及传播特征聚集在一起,形成各自的传播簇和流行簇。其中3对夫妻的毒株亚型和4对已知同性性伴侣的毒株亚型均为两两相同亚型,均分布于同一分支簇末端。
302例样本中共检出51例具有潜在(P)的≥1个的耐药突变位点,潜在突变率为16.89%(51/302),其中非核苷类反转录酶抑制剂( non-nucleoside reverse transcriptase inhibitors,NNRTIs)、核苷类反转录酶抑制剂(nucleoside reverse transcriptase inhibitors, NRTIs)、蛋白酶抑制剂(protein inhibitors,PIs)分别占比14.24%(43/302)、1.32%(4/302)、1.32%(4/302)。18例带有≥1个的低度至中高度耐药相关突变,耐药率为5.96%(18/302), NNRTIs、NRTIs、PIs分别占比4.97%(15/302)、0.66%(2/302)、0.33%(1/302),其中8例出现了低度耐药,10例出现了中高度耐药,但中高度耐药只出现于NNRTIs和PIs中,以依非韦伦(efavirenz,EFV)、奈韦拉平(nevirapine,NVP)尤为突出。
302例样本中,18例带有≥1个的低度至中高度耐药相关突变的HIV毒株亚型分布包括:CRF01_AE耐药率为3.67%(4/109)、CRF07_BC为4.92%(6/122)、URF为10.00%(2/20)、CRF55_01B为12.00%(3/25)、B为12.50%(1/8)、CRF08_BC为33.33%(2/6),不同亚型之间的耐药率差异有统计学意义(χ2=10.511,P=0.037);但中高度耐药发生率由低到高依次为CRF01_AE(2.75%,3/109)、CRF07_BC(3.28%,4/122)、CRF55_01B(4.00%,1/25)、URF(10.00%,2/20)。51例具有潜在(P)的≥1个的耐药突变的HIV毒株亚型分布包括:CRF01_AE为7.34%(8/109)、CRF07_BC为9.02%(11/122)、B为12.50 %(1/8)、CRF08_BC为16.67%(1/6)、URF为20%(4/20)、CRF67_01B为25%(1/4)、CRF55_01B 为88%(22/25),不同亚型之间的潜在耐药位点突变发生率差异有统计学意义(χ2=75.753,P<0.001)。其中CRF55_01B100%发生了耐药位点的突变。
本次调查结果显示,浦东新区感染HIV-1中青年人群虽然学历相对较高,且近年来各级疾控中心及基层社区卫生服务中心均推出了VCT服务以及大力的宣传教育,但早期发现以及主动检测的比例仍低于半数,且晚期发现占比较大,推测该人群尤其是异性性传播人群虽然清楚自己的高危性行为,但由于急性期无特异性或无任何不适症状,回避或侥幸心理成为了未及时发现的主要原因;还有部分同性性传播人群不认同口交或固定性伴侣或偶尔不戴套也会存在传播风险,从不主动检测,因未及时发现而成为感染HIV的源头,而性伴不固定以及跨区域、跨性别(同性及双性)等危险行为是HIV在中青年人群中传播的重要因素。
本研究共检测出11种亚型及多例独特重组型,说明浦东新区中青年HIV感染者人群中多种病毒亚型共存,呈现多元化流行趋势;其中CRF07_BC和CRF01_AE 为该人群优势毒株,这与上海市虹口区、浙江省湖州市及福建省的研究结果相近[10–12];同时结果显示,流行毒株CRF07_BC、CRF01_AE、CRF55_01B在15~24、25~34、35~49岁3个年龄段之间分布比较均衡,表明该人群HIV的性传播不仅跨越了地域界限同时也跨越了年龄界限。而CRF07_BC占比要高于CRF01_AE和CRF55_01B,推测一方面由于其同性和异性性传播的比例处于一个更加合适的比例范围,另一方面是由于同时具有同性和婚外异性性行为的比例尽管没有显著差异但还是高于其余2个亚型。研究显示B、C、CRF08_BC在该人群中未形成流行簇,推测一方面与各自亚型同性和异性性传播的比例未处在一个合适的比例范围,另一方面与年龄偏大尤其是异性性传播者以线下方式交友为主有较大关系。
本研究人群以同性性传播为主(72.19%),未婚、高学历、外地户籍占比较高,与中老年人群的人口学特征不同,但CRF07_BC、CRF01_AE 2种优势毒株仍与中老年感染人群非常接近[13],推测中青年人群与中老年人群有着较多的交集,并通过中老年人群向本地区其他人群扩散传播[14]。本次调查人群的传播耐药率处于中度流行水平,其中NNRTI耐药率最高;重组亚型CRF55_01BC 100%发生了耐药位点的突变,造成了88%的潜在耐药突变发生率及12%的耐药发生率,且存在中高度耐药,需要高度警惕并持续深入研究。目前国内各地报道的耐药位点也均以NNRTIs为主,同时NRTI及PIs的耐药位点并存[15–16]。据 WHO 相关报道,全球 PDR 的主要原因为NNRTIs耐药 [4],与本研究结果一致。
由于本研究是在浦东新区中老年HIV感染者分子流行病学分析基础上的补充,设计时考虑到中老年患者与>30岁组人群交集较多,所以<30岁人群的样本收集相对较少,会给结果带来一定的偏差,如感染平均年龄要比实际偏高、未婚率偏低等。但本文按照年龄段分层进行了流行毒株的特征分析,避免了其分子流行病学特征因所收集的样本由于年龄段占比不均衡带来的偏差。
综上,浦东新区中青年HIV感染人群中的病毒亚型呈现越来越多元化趋势,耐药突变位点发生率及不同程度的耐药率已出现于多种重组毒株中,因此应进一步加强中青年人群的干预力度,及时掌握高危动态,提高高危意识及早期发现的内驱力,以及持续加强耐药监测并给予抗病毒治疗前的指导服务。

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doi: 10.11847/zgggws1143438
  • 接收时间:2023-10-30
  • 首发时间:2026-10-25
  • 出版时间:2025-02-10
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  • 收稿日期:2023-10-30
  • 修回日期:2024-12-03
  • 录用日期:2025-01-26
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    1上海市浦东新区疾病预防控制中心,上海 2001362
    2复旦大学浦东预防医学研究院,上海 200136
    3苏州大学附属儿童医院,江苏 215025

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郝莉鹏,E-mail:
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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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