目的 了解青岛市男男性行为者(men who have sex with men,MSM)新发HIV-1感染者的病毒基因分型特征,构建HIV-1分子传播网络,为精准干预提供依据.方法 对2022年青岛市MSM新发HIV-1感染者进行问卷调查并采集血样,提取HIV-1病毒RNA,巢式PCR扩增pol区基因片段并进行测序,确定基因分型,计算基因距离,使用MEGA v11软件、HIV-Trace等构建系统进化树与分子网络,使用SPSS 27.0进行入网影响因素分析.结果 共调查158名MSM,采集152人血液样本,成功扩增135条pol区序列,共11种基因分型,主要基因分型为:CRF07_BC占42.23％(57/135),CRF01_AE 占 35.56％(48/135),CRF55_01B 占 9.63％(13/135);检出 了较为特殊的复杂重组体CRF01_BC和二代重组体CRF0107.鉴定出 13个传播簇,CRF07_BC形成最大的传播簇,包含43个节点,CRF01_AE包含15个节点,CRF55_01B包含11个节点.在0.03％的基因距离阈值下构建分子网络,入网率为51.11％(69/135).经x2检验与Fisher精确检验显示,HIV-1基因分型(x2=44.691,P＜0.01)、是否独居(x2=4.972,P=0.026)、对艾滋病了解程度(P=0.005)进入分子网络的差异有统计学意义.检测出44条序列存在耐药突变位点,耐药突变率为32.59％(44/135),21例进入分子网络.在基因型为CRF55_01B的一个传播簇中,所有感染者均对NNRTI耐药,耐药位点均为V179E.结论 青岛市MSM新发HIV-1感染者的病毒基因分型复杂多样,存在性活跃人群,尤其要把分型为CRF07_BC、非独居、对传染艾滋病抱有侥幸心理的MSM作为教育的重点,及时阻断传播链,避免分子网络的进一步扩大.
Genotyping and molecular network characteristics of newly reported HIV-1 infected patients among MSM in Qingdao
Objective To understand the genotyping characteristics of newly reported HIV-1 infected patients among MSM,and to construct a molecular transmission network of HIV-1,so as to provide accurate intervention for AIDS epidemic prevention and control in Qingdao.Methods A questionnaire survey was conducted in Qingdao for newly reported HIV-1 infected patients among MSM in 2022,and blood samples were collected to extract HIV-1 virus RNA.The gene fragments in pol region were amplified by nested PCR and sequenced for determination of sub-genotype and calculation of genetic distance.Phylogenetic trees and molecular networks were constructed by MEGA v11 and HIV-Trace software.SPSS 27.0 was used to analyze the influencing factors of accessing the network.Results A total of 158 MSM were investigated,and 152 blood samples were collected.One hundred and thirty-five pol region sequences were successfully amplified.There were 11 gene subtypes.For the major subtypes,CRF07_BC accounted for 42.23％(57/135),CRF01_AE for 35.56％(48/135)and CRF55_01B as 9.63％(13/135).Special complex recombinant strain CRF01_BC and second-generation recombinant strain CRF0107 were detected.Thirteen transmission clusters were identified,and CRF07_BC formed the largest cluster with 43 nodes while CRF01AE cluster had 15nodes and CRF55_01Bhad 11 nodes.The molecular network was constructed under the gene distance threshold of 0.03％,and the network access rate was 51.11％(69/135).According to thex2 test and Fisher's exact test,the differences of entering the molecular network caused by HIV-1 gene subtype(x2=44.691,P＜0.01),whether living alone(x2=4.972,P=0.026)and the understanding of AIDS(P=0.005)were all statistically significant.Drug-resistant mutation sites were detected in 44 sequences,with a drug-resistant mutation rate of 32.59％(44/135)Twenty-one cases entered the molecular network.In a transmission cluster of genotype CRF55_01B,all the infected people were resistant to NNRTI,and the resistant mutations were all V179E.Conclusions The subtypes of newly reported HIV-1 infected patients among MSM are complex and diverse,and there are sexually active people in MSM in Qingdao.Especially,Education for MSM population who were infected by subtype CRF07_BC,not living alone and had wishful thoughts of HIV infection should be focused to block transmission and further expansion of the molecular transmission network.
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