目的对保定市新发现的1株pol区不能明确分型的HIV-1毒株(BD226AJ)进行近似全长基因组扩增,并分析其亚型、重组模式和基因特点。方法提取患者血浆中HIV-1RNA并逆转录为cDNA,使用近末端稀释法分2段对其进行近似全长基因组扩增并测序。使用...目的对保定市新发现的1株pol区不能明确分型的HIV-1毒株(BD226AJ)进行近似全长基因组扩增,并分析其亚型、重组模式和基因特点。方法提取患者血浆中HIV-1RNA并逆转录为cDNA,使用近末端稀释法分2段对其进行近似全长基因组扩增并测序。使用jpHMM和SimPlot 3.5软件对近似全长基因组序列进行重组模式和重组断点分析,采用MEGA 6.0软件分片段构建Neighbor-joining系统进化树进一步确认重组断点的准确性。构建该近似全长基因组序列及各亚型片段Neighbor-joining系统进化树,分析该毒株的亲本来源。结果经过近似全长基因组扩增、测序、序列拼接后,获得1条长度为8830 bp的HIV-1近似全长基因组序列。重组分析结果显示,该序列是由CRF01_AE和B亚型重组形成的,其近似全长基因组序列被3个断点分成了4个亚型片段,分别为ICRF01_AE(HXB2,823—4224 nt)、Ⅱ_(B)(HXB2,4225—5991 nt)、ⅢCRF01_AE(HX B2,5992—9295 nt)、ⅣB(HXB2,9296—9406 nt)。各亚型基因片段的系统进化树分析进一步表明该序列的可能亲本来源为CRF01_AE和B亚型。HIV BLAST的结果显示,该序列与CRF112_01B的相似性为96%,系统进化树分析进一步验证该序列与北京市男男性行为者(men who have sex with men,MSM)中的CRF112_01B序列聚集。结论本研究在保定市MSM人群中发现了1例由CRF01_AE和B亚型重组的新型重组毒株CRF112_01B,提示HIV-1CRF112_01B已通过MSM人群传入河北,并开始在保定市传播,因此加强该亚型或者类似新型毒株的监测,对有关部门采取针对性防控措施和遏制新型重组毒株在本地区的传播和流行具有重要意义。展开更多
目的描述一株新鉴定的流行重组型HIV-1 CRF103_01B在北京的检出情况,并分析其近全长基因组(near full-length genome,NFLG)遗传特征。方法对2017—2020年北京新诊断或初始抗病毒治疗病例进行HIV-1基因型耐药监测,发现5例疑似感染CRF103_...目的描述一株新鉴定的流行重组型HIV-1 CRF103_01B在北京的检出情况,并分析其近全长基因组(near full-length genome,NFLG)遗传特征。方法对2017—2020年北京新诊断或初始抗病毒治疗病例进行HIV-1基因型耐药监测,发现5例疑似感染CRF103_01B毒株。通过逆转录、近末端稀释法,分两段巢式PCR扩增HIV-1 NFLG。获得的序列与分型参考序列进行基因比对,使用MEGA11软件构建邻接法(neighbor-joining,NJ)系统进化树。用SimPlot 3.5软件分析确定基因重组断点,绘制基因组结构图。基于亲本毒株同源NFLG序列比对,挑选较长的重组片段构建NJ进化树,判断可能的亲本毒株来源。用斯坦福大学HIV耐药数据库解析基因型耐药特征。结果共获得5条CRF103_01B NFLG序列,其中4例经男男性行为(men who have sex with men,MSM)感染,1例为女性。与从河北检出的4条CRF103_01B NFLG序列合并分析其重组特征:在CRF01_AE骨架上,gag、pol和nef-3'-LTR基因片段被B亚型重组替换[HXB2 nt 1111±19-1539±16,2531-4478±16(片段Ⅴ),9008±23-9615]。亲本来源分析显示,CRF103_01B的片段Ⅳ与Ⅴ分别与北京B亚型(BJMP3294B)和g5簇CRF01_AE序列(JX112804)聚集成大单系进化簇(bootstrap值分别为97%和100%)。9个CRF103_01B病例均携带非核苷类逆转录酶抑制剂类V106I突变。结论CRF103_01B毒株最可能起源于北京MSM人群,并在北京和河北等地的MSM人群中低水平持续流行,并经异性性途径向一般人群播散。需加强该毒株分子流行病学和耐药监测,关注疾病进展。展开更多
Laos is the only landlocked country in Southeast Asia and borders Thailand,Myanmar and Cambodia,the three countries in this region that have been hardest hit by human immunodeficiency virus(HIV).Laos has been regarded...Laos is the only landlocked country in Southeast Asia and borders Thailand,Myanmar and Cambodia,the three countries in this region that have been hardest hit by human immunodeficiency virus(HIV).Laos has been regarded as a low-HIVprevalence country for decades.To understand the status of HIV in Laos in recent years,a retrospective study was performed among 2851 patients visiting a hospital in Vientiane,the capital of Laos,from November 2011 to May 2012.Whole blood samples were obtained from the patients,and DNA was extracted.HIV status was determined by HIV gag fragment-specific PCR assay.Sixty-nine samples were detected as HIV proviral DNA positive with a positive rate of 2.4%(69/2851).Sixty-one near full-length genomic sequences were obtained from the positive samples.The results of phylogenetic analysis showed that the vast majority(91.8%)of the HIV strains belonged to CRF01AE,and the other five(8.2%)strains were identified as a new HIV circulating recombinant form CRF9701B,which had a CRF01AE backbone with an insertion of subtype B in the gag-pol region.Phylogeographic analysis revealed that HIV CRF01AE circulating in Laos were multiply introduced from Thailand.These results indicated that Laos might be suffering a considerably more serious impact of HIV than previously believed.To keep this country from undergoing the same increase in HIV prevalence observed in its neighbors,immediate intervention measures and sufficient epidemiological research are urgently needed.展开更多
文摘目的对保定市新发现的1株pol区不能明确分型的HIV-1毒株(BD226AJ)进行近似全长基因组扩增,并分析其亚型、重组模式和基因特点。方法提取患者血浆中HIV-1RNA并逆转录为cDNA,使用近末端稀释法分2段对其进行近似全长基因组扩增并测序。使用jpHMM和SimPlot 3.5软件对近似全长基因组序列进行重组模式和重组断点分析,采用MEGA 6.0软件分片段构建Neighbor-joining系统进化树进一步确认重组断点的准确性。构建该近似全长基因组序列及各亚型片段Neighbor-joining系统进化树,分析该毒株的亲本来源。结果经过近似全长基因组扩增、测序、序列拼接后,获得1条长度为8830 bp的HIV-1近似全长基因组序列。重组分析结果显示,该序列是由CRF01_AE和B亚型重组形成的,其近似全长基因组序列被3个断点分成了4个亚型片段,分别为ICRF01_AE(HXB2,823—4224 nt)、Ⅱ_(B)(HXB2,4225—5991 nt)、ⅢCRF01_AE(HX B2,5992—9295 nt)、ⅣB(HXB2,9296—9406 nt)。各亚型基因片段的系统进化树分析进一步表明该序列的可能亲本来源为CRF01_AE和B亚型。HIV BLAST的结果显示,该序列与CRF112_01B的相似性为96%,系统进化树分析进一步验证该序列与北京市男男性行为者(men who have sex with men,MSM)中的CRF112_01B序列聚集。结论本研究在保定市MSM人群中发现了1例由CRF01_AE和B亚型重组的新型重组毒株CRF112_01B,提示HIV-1CRF112_01B已通过MSM人群传入河北,并开始在保定市传播,因此加强该亚型或者类似新型毒株的监测,对有关部门采取针对性防控措施和遏制新型重组毒株在本地区的传播和流行具有重要意义。
文摘目的描述一株新鉴定的流行重组型HIV-1 CRF103_01B在北京的检出情况,并分析其近全长基因组(near full-length genome,NFLG)遗传特征。方法对2017—2020年北京新诊断或初始抗病毒治疗病例进行HIV-1基因型耐药监测,发现5例疑似感染CRF103_01B毒株。通过逆转录、近末端稀释法,分两段巢式PCR扩增HIV-1 NFLG。获得的序列与分型参考序列进行基因比对,使用MEGA11软件构建邻接法(neighbor-joining,NJ)系统进化树。用SimPlot 3.5软件分析确定基因重组断点,绘制基因组结构图。基于亲本毒株同源NFLG序列比对,挑选较长的重组片段构建NJ进化树,判断可能的亲本毒株来源。用斯坦福大学HIV耐药数据库解析基因型耐药特征。结果共获得5条CRF103_01B NFLG序列,其中4例经男男性行为(men who have sex with men,MSM)感染,1例为女性。与从河北检出的4条CRF103_01B NFLG序列合并分析其重组特征:在CRF01_AE骨架上,gag、pol和nef-3'-LTR基因片段被B亚型重组替换[HXB2 nt 1111±19-1539±16,2531-4478±16(片段Ⅴ),9008±23-9615]。亲本来源分析显示,CRF103_01B的片段Ⅳ与Ⅴ分别与北京B亚型(BJMP3294B)和g5簇CRF01_AE序列(JX112804)聚集成大单系进化簇(bootstrap值分别为97%和100%)。9个CRF103_01B病例均携带非核苷类逆转录酶抑制剂类V106I突变。结论CRF103_01B毒株最可能起源于北京MSM人群,并在北京和河北等地的MSM人群中低水平持续流行,并经异性性途径向一般人群播散。需加强该毒株分子流行病学和耐药监测,关注疾病进展。
基金supported in part by Grants from the National Natural Science Foundation of China(U1302224,81601802,81271892,81902106)
文摘Laos is the only landlocked country in Southeast Asia and borders Thailand,Myanmar and Cambodia,the three countries in this region that have been hardest hit by human immunodeficiency virus(HIV).Laos has been regarded as a low-HIVprevalence country for decades.To understand the status of HIV in Laos in recent years,a retrospective study was performed among 2851 patients visiting a hospital in Vientiane,the capital of Laos,from November 2011 to May 2012.Whole blood samples were obtained from the patients,and DNA was extracted.HIV status was determined by HIV gag fragment-specific PCR assay.Sixty-nine samples were detected as HIV proviral DNA positive with a positive rate of 2.4%(69/2851).Sixty-one near full-length genomic sequences were obtained from the positive samples.The results of phylogenetic analysis showed that the vast majority(91.8%)of the HIV strains belonged to CRF01AE,and the other five(8.2%)strains were identified as a new HIV circulating recombinant form CRF9701B,which had a CRF01AE backbone with an insertion of subtype B in the gag-pol region.Phylogeographic analysis revealed that HIV CRF01AE circulating in Laos were multiply introduced from Thailand.These results indicated that Laos might be suffering a considerably more serious impact of HIV than previously believed.To keep this country from undergoing the same increase in HIV prevalence observed in its neighbors,immediate intervention measures and sufficient epidemiological research are urgently needed.