2018年至2019年浙江省绍兴市新确证人类免疫缺陷病毒1型感染者的分子传播网络特征分析

Characteristics of the molecular transmission network in newly confirmed human immunodeficiency virus type 1 infected cases from 2018 to 2019 in Shaoxing City,Zhejiang Province

目的分析浙江省绍兴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。方法纳入2018年8月至2019年12月绍兴市新确证未经抗病毒治疗的423份HIV-1感染/艾滋病病例血液样本,获得375份样本序列,采用反转录聚合酶链反应和巢式聚合酶链反应扩增HIV-1的pol基因,采用MEGA 6.0软件构建系统进化树分析亚型,并采用HyPhy软件和Cytoscape 3.7.2。生成不同基因距离的分子网络,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375份样本序列中发现8种亚型,优势亚型为流行重组型(circulating recombinant form,CRF)07BC 215份(57.33%),CRF01AE 103份(27.47%),其他包括CRF08BC、CRF85BC、CRF5501B、B、C和CRF01AE/B亚型。在基因距离为1.50%时,共形成24个分子簇,194条序列入网,入网率为51.73%。在分子簇最多的0.75%基因距离时,共形成30个分子簇,129条序列入网,入网率为34.40%,35例以老年人为主的病例聚集成CL1簇。42份样本存在监测性耐药突变(surveillance drug resistance mutation,SDRM),耐药传播率为11.20%(42/375),其中非核苷类反转录酶抑制剂耐药突变38例,分别为K103N(32/375,8.53%)、K103S(4/375,1.07%)、Y188L(1/375,0.27%)和G190A(1/375,0.27%),蛋白酶抑制剂耐药突变4例,分别为M46I(3/375,0.80%)和V82A(1/375,0.27%)。分子簇C2序列携带高比例耐药突变(94.29%,33/35)。结论绍兴市HIV-1亚型丰富多样,CRF07BC亚型传播迅速;在0.75%基因距离仍聚集的CL1簇老年病例亟待干预,防止该耐药毒株的进一步快速传播。

Objective To analyze the characteristics of the molecular transmission network of newly-diagnosed human immunodeficiency virus type 1(HIV-1)infected individuals in Shaoxing City,Zhejiang Province,and to provide evidence for epidemic trend and prevention.Methods The plasma samples from 423 antiretroviral-naive HIV-1/acquired immunodeficiency syndrome patients from August 2018 to December 2019 were collected,and the pol gene fragments of HIV-1 from 375 samples were amplified by reverse transcription polymerase chain reaction(PCR)and nested PCR.The phylogenetic tree was constructed to analyze the molecular transmission network for subtypes and different gene distances by MEGA 6.0 software,HyPhy software and Cytoscape 3.7.2.Mutations on drug resistance was analyzed by online software tool of the HIV drug resistance database of Stanford University.Results Eight subtypes were found in the 375 samples.Circulating recombinant form(CRF)07_BC(215/375,57.33%)and CRF01_AE(103/375,27.47%)were the major subtypes,followed by CRF85_BC,CRF55_01B,B,C,and CRF01_AE/B subtypes.One hundred and ninety-four individuals(51.73%)were connected to the transmission network at 1.50%genetic distance with 24 clusters.One hundred and twenty-nine individuals(34.40%)were connected to the transmission network at 0.75%genetic distance with 30 clusters,and 35 elderly patients were clustered in CL1.Forty-two cases had surveillance drug resistance mutation(SDRM),the prevalence of transmitted drug resistance was 11.20%(42/375).Thirty-eight cases had the drug mutations to non-nucleoside reverse transcriptase inhibitor,including K103N(32/375,8.53%),K103S(4/375,1.07%),Y188L(1/375,0.27%)and G190A(1/375,0.27%);four cases had the mutations to protease inhibitor,including M46I(3/375,0.80%)and V82A(1/375,0.27%).The cluster C2 sequences carried a high proportion of resistant mutations(94.29%,33/35).Conclusions HIV-1 subtypes in Shaoxing City are diverse and the CRF07_BC subtype spreads rapidly.The elderly patients with drug resistance genes in cluster CL1 at 0.75%gene distance need to be intervened immediately to prevent the drug resistance virus spread.