我国西南西北地区吸毒人群重组人类免疫缺陷病毒1？…

目的 寻找人类免疫缺陷病毒１型在中国可能的重组。方法从流行２种以上ＨＩＶ－１亚型的地区收集ＨＩＶ感染者的血样。从ＰＭＣｓ中应用套式聚合酶链反应方法,对ＨＩＶ病毒的ｔａｔ和ｅｎｖ基因进行扩增,ＰＣＲ产物直接测序并进行序列分析。结果 对中国Ｂ’亚型和Ｃ亚型浒区域收集的１４个ＨＩＶ－１毒株进行序列分析,对ｅｎｖ基因进行测序后没有发现重组毒株的证据。     

中国人类免疫缺陷病毒（HIV—1）D亚型毒株gag,env和t …

目的 通过对ＨＩＶ－１毒株ｇａｇ,ｔａｔ,ｅｎｖ基因的序列分析,阐明Ｄ亚型ＨＩＶ－１毒株已在中国出现。方法 从１名四川非洲回国营务人员ＨＩＶ感染者（ＳＣ９ ７１２）淋巴细胞（ＰＢＭＣ）中提取前病毒ＤＮＡ,使用套式ＰＣＲ方法分别扩增ＨＩＶ－１的ｇａｇ基因区,ｅｎｖ基因的Ｃ２Ｖ３区和ｔａｔ基因的第一外显子。结果 发现ＳＣ９ ７１２在ｇａｇ区,ｅｎｖ区和ｔａｔ区与国际标准Ｄ亚型毒株的基因距离最近,其中在     

山东省某些高危人群HIV-1感染者分子流行病学研究

目的 了解流行于山东境内ＨＩＶ－１毒株的亚型分布及变异情况,分析其来源并推测其流行趋势。方法 采集了２５份ＨＩＶ－１抗体阳性感染者的全血分离单核细胞（ＰＢＭＣ）,提取前病毒ＤＮＡ,经ｎｅｓｔｅｄ－ＰＣＲ扩增ＨＩＶ－１膜蛋白（ｅｎｖ）基因的Ｃ２－Ｖ３区并进行序列测定和亚型分析。结果 ２５例ＨＩＶ－１阳性感染者的ＰＢＭＣ样品中扩增到２４份可用于序列测定的ＨＩＶ－１ｅｎｖ基因片段,经序列测定和基因分析鉴     

云南省陇川县HIV-1流行毒株膜蛋白基因C2-V3区序列测定和分析

使用ＰＣＲ对２１份采集于１９９５年中的云南陇川县ＨＩＶ－１阳性静脉吸毒者外周血单个核细胞（ＰＢＭＣｓ）样品进行扩增，从１７份样品中获得了ＨＩＶ－１膜蛋白（ｅｎｖ）基因的核酸片段，并对其Ｃ２－Ｖ３及邻区４５０个核苷酸序列进行了测定和分析。研究结果表明，１７份陇川样品中存在Ｂ和Ｃ两种亚型的ＨＩＶ－１毒株序列，各亚型内的基因离散率分别为４．７％和３．３％。与Ａ～Ｅ参考亚型及部分Ｂ和Ｃ亚型代表株序列相比较，属陇川Ｂ亚型的１４个毒株与包括泰国、缅甸及云南瑞丽在内的Ｂ亚型毒株序列十分接近，基因离散率在３．９％～４．５％的范围内；属陇川Ｃ亚型的３个毒株则与代表印度毒株的Ｃ亚型共享序列及瑞丽Ｃ亚型毒株序列十分相似，其基因离散率均为２．５％。以上数据提示，ＨＩＶ－１在陇川的流行时间不长，且Ｂ和Ｃ亚型毒株的传入时间相差１年左右。对Ｂ亚型毒株Ｖ３环序列的分析还发现，位于Ｖ３环顶端的四肽序列中ＧＰＧＱ占６４．３％，ＧＰＧＲ则仅占２８．６％，且编码其精氨酸（Ｒ）的密码子均为ＣＧＡ而不是ＡＧＡ。此结果与作者根据早期瑞丽ＨＩＶ－１毒株序列研究结果得出的推测相吻合。     

克隆中国B,C,E亚型的HIV—1代表株建立适于我国流行株的 …

目的 构建我国人免疫缺陷病毒１型（ＨＩＶ－１）Ｂ、Ｃ和Ｅ各亚型代表株ｅｎｖ基因质粒,用于对中国ＨＩＶ－１进行分型。方法 将与我国ＨＩＶ－１ Ｂ,Ｃ和Ｅ各亚型共享序列最接近的毒株用套式聚合酶链反应（ＰＣＲ）技术扩增包膜（ｅｎｖ）基因,克隆到ｐＧＥＭ－Ｔｅａｓｙ载体中,构建成用于异源双链泳动分析（ＨＭＡ）分型的中国标准亚型质粒,并进行了序列分析和分型的敏感性分析。结果 构建的中国ＨＩＶ－１ Ｂ,Ｃ和Ｅ     

云南省陇川县HIV—1流行毒株膜蛋白基因C2—V3区序列测定…

使用ＰＣＲ对２１份采集于１９９５年中的云南陇川县ＨＩＶ－１阳性静脉吸毒者外周血单个核细胞（ＰＢＭＣｓ）样品进行扩增，从１７份样品中获得了ＨＩＶ－１膜蛋白（ｅｎｖ）基因的核酸片段，并对其Ｃ２－Ｖ３及邻区４５０个核苷酸序列进行了测定和分析。研究结果表明，１７份陇川样品中存在Ｂ和Ｃ两种亚型的ＨＩＶ－１毒株序列，各亚型内的基因离散率分别为４．７％和３．３％。与Ａ￣Ｅ参考亚型及部位Ｂ和Ｃ亚型代表株序列相比较     

深圳市HIV-1 E亚型感染毒株的分子流行病学分析

目的 了解人类免疫缺陷病毒１型Ｅ亚型毒株在深圳市不同人群中流行传播情况、流行时间和传播规律。方法 应用聚合酶链反应（ＰＣＲ）方法对１９９６年深圳市检出的３份人类免疫缺陷病毒１型（ＨＩＶ－１）感染者外周血单个核细胞样本进行扩增,获得ＨＩＶ－１膜蛋白（ｅｎｖ）基因片段,并对Ｃ２－Ｖ３及其邻区３５０～４５０个核苷酸序列进行测定和分析。结果 这３份血样为ＨＩＶ－１Ｅ亚型毒株感染（ｓｚ－Ｅ）,彼此间的基因离散率为２．６％;与Ａ－Ｅ国际参考亚型及国内部分地区流行的ＢＥ型代表株比较,ｓｚ－Ｅ与Ａ－Ｄ参考亚型共享序列及国内Ｂ亚型代表株间的基因离散率均大于２４％,而与主要代表泰国Ｅ亚型（Ｅｃｏｎ）间的基因离散率仅为６．２％。系统树分析显示,ｓｚ－Ｅ与Ｅｃｏｎ聚集在一起,远离其他国际亚型毒株序列。结论 ＨＩＶ－１Ｅ亚型在深圳的流行     

我国云南省艾滋病病毒感染的长期不进展者人免疫缺陷 …

目的 人类免疫缺陷病毒１型（ＨＩＶ－１）ｔａｔ基因是该病毒的调控基因之一。本研究是探讨ｔａｔ基因变异是否影响ＨＩＶ－１感染者的病程进展。方法 从云南ＨＩＶ流行区的２２例感染后临床进程不同的人抽取外周血,提取核酸,用套式聚合酶链反应（ＰＣＲ）扩增ＨＩＶ－１的ｔａｔ基因,并进行了核酸序列测定和分析。     

丙型肝炎病毒非结构基因5b(NS 5b)区一级结构的变异

目的　分析中国丙型肝炎病毒非结构基因５ ｂ 区核苷酸序列变异。方法　以逆转录套式聚合酶链反应（ＲＴｎｅｓｔｅｄＰＣＲ） 从４９ 例中国病人的血清中获得互补的ＤＮＡ片段,产物克隆后测序。结果　３３ 株系基因Ⅱ１ ｂ 型,１６ 株系Ⅲ２ ａ 型,中国ＨＣＶ株型与日本ＨＣＶ株型同属１ 个基因亚型,但在一些核苷酸保守位点上有一定的差异,对３３ 株Ⅱ１ ｂ 型和１６ 株Ⅲ２ ａ 型间的同源性进行比较,发现１６ 株Ⅲ２ ａ 型的同源性低于３３ 株Ⅱ１ ｂ 型的同源性。首次在ＨＣＶＮＳ５ ｂ 区发现１ 个新的３ 个核苷酸的缺失突变及１ 个移码突变。结论　同一基因亚型之内不同ＨＣＶ株的核苷酸序列可能具有一定的地理分布特征,每个地区有一定的流行株。     

一株HIV1变异株的发现

我们在进行序列分析过程中发现一株不同于以往国内外报道的新型ＨＩＶ1的变异毒株。现报道如下 :1　材料和方法1.1　前病毒ＤＮＡ的提取　某ＨＩＶ1感染者 30 0 μｌＥＤＴＡ抗凝血 ,使用Ｐｒｏｍｅｇａ公司的ＤＮＡ提取试剂盒 (ＷｉｚａｒｄＧｅｎｏｍｉｃＤＮＡＰｕｒｉｆｉｃａｔｉｏｎＫｉｔ)提取ＨＩＶ1前病毒ＤＮＡ。1.2　ＨＩＶｅｎｖ基因ｃ2 -ｖ3区的套式ＰＣＲ扩增　以所提的ＨＩＶ1前病毒ＤＮＡ为模板 ,利用一对外侧引物ＥＰ1,ＥＰ2 (序列为 ,ＥＰ1:5′ -ＧＧＣＡＧＴＣＴＡＧＣＡ ＧＡＡＧＡＡＧＡＧＧ - 3′ ;ＥＰ2 :5′ -ＣＣ…     

Subtype CRF01_AE dominate the sexually transmitted human immunodeficiency virus type 1 epidemic in Guangxi,China

Human immunodeficiency virus type 1 (HIV‐1) infection by sexual transmission in Guangxi, China had increased dramatically. However, limited information is available on the genetic characterization of the HIV‐1 epidemic. In this study, HIV‐1 seropositive drug‐naïve patients infected by heterosexual transmission were enrolled. The full length gag and pol genes were sequenced followed by phylogenetic analysis, recombinant analysis and drug resistant analysis. Multiple subtypes were identified, including CRF01_AE (80.1%), CRF07_BC (6.4%), CRF08_BC (10.2%), subtype B (1.7%), and URFs (1.7%). In the phylogenetic tree, two large CRF01_AE clusters were identified. One cluster is originating from Vietnam strains as being reported previously in intravenous drug users. One novel cluster was identified and showed close relationship to strains from Fujian province. Inter‐subtype recombination among CRF01_AE, subtype B and C was identified. Low level drug‐resistance in drug‐naïve heterosexually transmitted infections was found. The results suggested that multiple originating CRF01_AE strains dominated the HIV‐1 epidemic in heterosexual transmission in Guangxi province. J. Med. Virol. 85:388–395, 2013. © 2013 Wiley Periodicals, Inc.     

Determination of human immunodeficiency virus type 1 subtypes in Taiwan by vpu gene analysis

The genetic diversity of human immunodeficiency virus (HIV) type 1 (HIV-1) has been characterized mainly by analysis of the env and gag genes. Information on the vpu genes in the HIV sequence database is very limited. In the present study, the nucleotide sequences of the vpu genes were analyzed, and the genetic subtypes determined by analysis of the vpu gene were compared with those previously determined by analysis of the gag and env genes. The vpu genes were amplified by nested PCR of proviral DNA extracted from 363 HIV-1-infected individuals and were sequenced directly by use of the PCR products. HIV-1 subtypes were determined by sequence alignment and phylogenetic analysis with reference strains. The strains in all except one of the samples analyzed could be classified as subtype A, B, C, E, or G. The vpu subtype of one strain could not be determined. Of the strains analyzed, genetic subtypes of 247 (68.0%) were also determined by analysis of the env or gag gene. The genetic subtypes determined by vpu gene analysis were, in general, consistent with those determined by gag and/or env gene analysis except for those for two AG recombinant strains. All the strains that clustered with a Thailand subtype E strain in the vpu phylogenetic analyses were subtype E by env gene analysis and subtype A by gag gene analysis. In summary, our genetic typing revealed that subtype B strains, which constituted 73.8% of all strains analyzed, were most prevalent in Taiwan. While subtype E strains constituted about one-quarter of the viruses, they were prevalent at a higher proportion in the group infected by heterosexual transmission. Genetic analysis of vpu may provide an alternate method for determination of HIV-1 subtypes for most of the strains, excluding those in which intersubtype recombination has occurred.     

26例有偿献血员HIV-1基因分型与变异特征

目的分析某地区有偿献血人员中流行的人免疫缺陷病毒1型（HIV-1）gag、pol、env基因亚型及基因变异特征。方法提取HIV-1感染者外周血单核细胞（PBMC）DNA,经巢式PCR（NestedPCR）扩增gag（p17-p24）、pol（PR＊RT）、env（C2-V5）基因片段,纯化测序后用MEGA5．0等生物学软件对核苷酸序列进行分析。结果23份样本为B亚型,2份为B亚型与C亚型重组,1份为CRF01-AE与B亚型重组。PR区未发现蛋白酶抑制剂主要耐药性突变,RT区检测到核苷类逆转录酶抑制剂耐药性突变M184V和非核苷类逆转录酶抑制剂耐药性突变K101E,G190A。结论流行于该地区的HIV-1毒株以B亚型为主。大多数毒株对常规抗病毒药物仍然敏感,使用HARRT治疗方案依然有效。CXCR4型辅助受体的毒株顶端四肽多为GPGR（91．7％）,提示GPGR可能与疾病的进展有关。     

中国HIV-1型B、C亚型主要流行株外膜蛋白基因V3-V4区序列的变异分析

目的　研究我国人类免疫缺陷病毒 1型 (HIV 1)B、C亚型主要流行株在感染过程中基因变异的特点及其与选择压力的关系。方法　应用巢式聚合酶链反应 (nested PCR)对 2 5 8例HIV 1感染者血样中的HIV 1外膜蛋白 (env)基因进行扩增 ,并使用ABI 377型测序仪对扩增产物测序后 ,选择其中 37份B亚型和 35份C亚型HIV 1毒株env基因包括V3～V4区的序列进行比较分析 ,并计算和分析氨基酸同义替换与非同义替换的比值 (Ks Ka)。结果　B亚型毒株V3～V4区的基因离散率高于C亚型毒株。无论B亚型 ,还是C亚型毒株 ,其V4区基因序列较V3区变异更大。在C亚型毒株中 ,V3区基因序列变异甚至比V3上游区和C3区小。B和C亚型毒株整个V3～V4基因区的Ks Ka比值均 <1,差异有非常显著性 (P <0 0 0 1) ,其中B亚型毒株以V3区的Ks Ka比值最小 ,而C亚型毒株则以V4区的Ks Ka比值最小。结论　B和C亚型毒株env基因的变异主要发生在V4区而不是V3区。C亚型毒株V3区较V3上游区和C3区还要保守 ,是本研究的特殊发现。这两种亚型在我国快速流行中发生的变异是在选择压力下发生的 ,而不是随机进化的结果 ,而且选择压力对这两种亚型毒株V3、V4区的作用程度也不一样。这将为我国艾滋病防治策略的制定和疫苗研究提供科学的依据。     

Molecular epidemiology of HIV-1 in Switzerland: evidence for a silent mutation in the C2V3 region distinguishing intravenous drug users from homosexual men. Swiss HIV Cohort Study

OBJECTIVES: To study the molecular epidemiology of HIV-1 strains found in Switzerland and to determine possible genetic linkages among strains sorted by risk group or geographic region. DESIGN: A cross-sectional, clinic-based survey of HIV-1 molecular sequences and linked patient history from Swiss people. METHODS: Specimens were collected from 215 HIV-1-infected people in HIV outpatient clinics of four tertiary referral centers (Lausanne, St. Gallen, Zurich, and Basel) between May and August 1996, mainly from homosexual men, injecting drug users (IDU), and heterosexually infected people. In addition, specimens collected between 1991 and 1995 in the HIV outpatient clinic at University of Geneva were included into this survey. These specimens were collected primarily for an ongoing, prospective cohort (Swiss HIV Cohort Study). Direct C2V3C3 sequences of the env gene were determined from 158 samples of peripheral blood mononuclear cells. Genetic data were analyzed with the available patient history on each specimen. RESULTS: As found in other previous studies in Europe, primarily subtype B viruses were identified, whereas seven (4%) of 158 were non-subtype B: one subtype D, four subtype A, and two subtype E. Five of seven non-B subtypes occurred in immigrants from African or Asian countries and all seven were found exclusively in individuals who had been infected by heterosexual contact. No significant clustering of strains within different study sites or risk groups was found. A silent mutation (LAI env 834) occurred significantly more often in IDU than in homosexual men (p<.001). CONCLUSIONS: Although the lack of significant clustering of strains by risk group or geographic region may result from early introduction of subtype B viruses in Switzerland, the strong association of a silent mutation with IDU suggests that, early in the epidemic, there was a unique founder virus among IDUs. The HIV epidemic in Switzerland is still predominantly caused by subtype B viruses.     

Near full-length genomic characterization of a HIV type 1 BC recombinant strain from Manipur, India

Genetic complexity of HIV-1 is brought about by recombination between HIV-1 subtypes which leads to the development of epidemiologically significant founder strains. In the present study, the near full-length genome sequence of an HIV-1 isolate from an injecting drug user of Manipur (India) was determined, which evidenced the presence of a novel HIV-1 BC recombinant strain. Near full-length genome was amplified by polymerase chain reaction using primer walking approach. The recombination break points were detected using bootscan and simplot analyses. This isolate exhibited a mosaic structure consisting of subtype C backbone with subtype B insertions at the upstream of pol gene (3026-3259) and the downstream of env gene which spanned till the nef gene (8183-8961). Phylogenetic relationships determined with neighbor-joining trees, revealed that the subtype C sequences clustered with sequences from Indian subtype C HIV-1 strains, and the subtype B sequences clustered with HIV-1 subtype B strains from Thailand. This finding may create a complex scenario of HIV-1 epidemic among the injecting drug users of Manipur in near future.