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1.
应用异源双链泳动分析法快速确定(HIV-1)基因亚型   总被引:10,自引:0,他引:10  
应用异源双链泳动分析法(Heterouduplex mobility assay,HMA),对来自我国云南、四川、新疆、浙江、广东、福建、天津7省市73份HIV感染者样品,进行了HIV-1膜蛋白(env)基因片段亚型分析。通过HMA能确定亚的有70份,占样品总数的95.89%(70/72)。其中A亚型2.86%(2/70),F亚型2.86%(2/70),泰国B亚型18.58%(13/70),欧美B  相似文献   

2.
为了解HIV抗体阳性血浆中的HIV1病毒基因亚型的情况,应用逆转录PCR和DNA序列测定技术,对6份获自高危人群的抗HIV1阳性血浆进行序列分析和基因亚型分型的研究,结果表明均属HIV1B亚型。V3环氨基酸序列分析指出这些HIV1B亚型病毒株与泰国HIV1B亚型病毒株核苷酸和氨基酸序列相似;同时发现HIV1cDNA和氨基酸序列均相同,推测这6份标本可能来自同时感染同一株HIV病毒的感染者。本研究对了解高危人群中HIV1流行的遗传变异和HIV1亚型病毒株的分子流行病分析具有一定的意义。  相似文献   

3.
陈杰  苏玲中 《病毒学报》1998,14(3):240-245
使用PCR技术对14份广西HIV-1阳性感染者外周血单核细胞(PBMCs)样品进行扩增,获得HIV-1膜蛋白(env)基因的核酸片段,并对其C2-V3及邻区350 ̄450个核苷酸序列进行了测定和分析。结果表明,14份样品中9份为泰国B(B')亚型,5份为E亚型毒株。其中B'亚型毒株的基因离散率为4.2%,与A-E参考亚型及部分B亚型代表株序列相比较,与包括泰国、缅甸及云南德宏在内的B亚型毒株序列十  相似文献   

4.
广西壮族自治区HIV-1流行毒株的基因序列测定和亚型分析   总被引:12,自引:0,他引:12  
使用PCR技术对14份广西HIV-1阳性感染者外周血单核细胞(PBMCs)样品进行扩增,获得HIV-1膜蛋白(env)基因的核酸片段,并对其C2-V3及邻区350-450个核苷酸序列进行了测定和分析。结果表明,14份样品中9份为泰国B(B′)亚型,5份为E亚型毒株。其中B′亚型毒株的基因离散率为4.2%,与A-E参考亚型及部分B亚型代表株序列相比较,与包括泰国、缅甸及云南德宏在内的B亚型毒株序列十分接近,相互之间基因离散率在3.0%-4.4%的范围内;而E亚型毒株的基因离散率为2.1%,与国际E亚型毒株的基因离散率最近,为5.6%,与其它国际参考亚型基因离散率很远,在21.1%-27.3%。根据以上数据及其它资料提示,广西存在B′和E两种亚型的HIV-1的流行,且其B′亚型毒株的传入,与流行在云南德宏州的相同亚型HIV-1毒株密切相关,而E亚型毒株则可能是由泰国经越南传入广西的  相似文献   

5.
1995年云南瑞丽HIV1毒株的基因变异和分析   总被引:28,自引:3,他引:25  
从1995年瑞丽静脉注射毒品者及其配偶26人的血样中,经PCR扩增细胞内HIV基因并对其envC2-V3区进行测序,结果发现18人为B亚型HIV1,8人为C亚型HIV1毒株,在瑞丽以B亚型毒株为主,与我们以往报告一致,C亚型毒株的出现曾有报告,我们进一步分析发现,C亚型毒株感染者多在1994和1995年被感染,根据其核苷酸序列测定的基因离散率(2.25%),远小于B型毒株(6.7%),也说明它是近  相似文献   

6.
人源单克隆抗人免疫缺陷病毒1型抗体Fab段基因的获得   总被引:1,自引:0,他引:1  
应用噬苏体抗体库技术有效地筛选出了多株抗HIV-1人源单克隆抗体。以逆转录聚合酶链反应(RT-PCR)从HIV-1感染者外周血淋巴细胞中扩增抗体轻重链可变区基因,插入载体pCOMB3,建立噬菌体抗体库。分别以HIV-1gp120和gp160为固相抗原,经过多轮筛选,从中获得了多株抗HIV-1gp41、gp120和gp160的单克隆抗体Fab段基因。抗HIV特异性噬菌体抗体随抗体库的筛选高度富集,抗  相似文献   

7.
邢辉  戴玉琳 《病毒学报》2000,16(2):111-115
上海市卫生检疫局送检了一例HIV-1和HIV-2抗体检测均呈阳性的双重感染样品,对其感染的HIV前病毒的gag和env基因区进行了序列分析,首次阐明我国发现的HIV双重感染样品的HIV部分基因特征。从HIV感染者淋巴细胞(peripheral blood mononuclear cells,PBMC)中提取前病毒DNA,分别使用HIV-1和HIV-2特异性引特用套式PCR扩增HIV-1和HIV-2  相似文献   

8.
缺氧诱导因子1研究进展   总被引:3,自引:0,他引:3  
缺氧诱导因子1(HIF-1)在缺氧诱导的哺乳动物细胞中广泛表达,为缺氧应答的全局性调控因子。HIF-1由HIF-1α和HIF-1β两亚基组成,为异源二聚体转录因子。HIF-1α的bHLH和PAS结构域与二聚化及DNA结合活性有关,TAD结构域则主要参与转录激 活。HIF-1α的全长基因已克隆并在人和小鼠中定位。通过作用于靶基因的缺氧反应元件(HRE0,HIF-1参与缺氧诱导的一系列基因的表达调控。  相似文献   

9.
莫凌 《微生物与感染》1999,22(6):8-10,16
人类免疫缺陷病毒(HIV)载量是指HIV感染者体内潜伏的和具有复制活性的病毒数,它包括血浆中的循环HIV-RHA和整合到细胞内的HIV-DNA即前病毒。近年来随着检测技术的发展,人们发现早期感染者的组织中已存在大量病毒和病毒产物,因此可用病毒载量来评估无症状感染者的免疫状态和病毒的复制情况。近年来病毒载量的测定已被广泛地用于多项临床研究及观察中,现就HIV感染者病毒载量研究的若干进展作一简要综述。  相似文献   

10.
将中国株HIV-1B亚型gp120全基因序列克隆到杆状病毒转座载体pFastBacI中多角体启动子下游,构建成重组转座载体pFastBacI-gp120,利用细菌/杆状病毒(Bac to Bac)表达系统筛选重组杆状病毒,在昆虫细胞Sf9中高效表达了HIV-1的外膜糖蛋白gp120,SDS-PAGE和Western blot分析结果一致,证明表达了2种糖基化程度不同的gp120。  相似文献   

11.
DNA sequences encoding the C2 to V3 region of envelope glycoprotein gp120 of human immunodeficiency virus type 1 (HIV-1) were amplified by PCR from uncultured peripheral blood mononuclear cells obtained from 24 of 25 HIV-1-seropositive patients from Cyprus. By using a heteroduplex mobility assay (HMA), all amplified products were studied genetically and compared with 16 previously characterized HIV-1 strains belonging to subtypes A through F. HMA results revealed that HIV-1 gp120 sequences from 15 of our patients were of subtype B of HIV-1, whereas one isolate was of subtype C. However, gp120 sequences from eight patients had no obvious similarities to the known subtypes as defined by HMA. DNA sequencing and phylogenetic analyses of molecular clones confirmed the HMA results and placed the eight undefined HIV-1 isolates into three distinct genetic clusters. On the basis of branch topology and lengths of the phylogenetic tree, we conclude that one group consisting of three clones from two patients represents a new HIV-1 env subtype, which we have termed subtype I. The remaining two sequence clusters, consisting of five sequences from four patients and two sequences from two other patients, are distally related to subtypes A and F. These data demonstrate the extensive heterogeneity of HIV-1 in Cyprus, including the presence of new subtype.  相似文献   

12.
Genetic variability of human immunodeficiency virus type-1(HIV-1) is a potential threat for both diagnosis and treatment of HIV/AIDS, as well as the development of effective vaccines. Up to now, HIV subtypes circulating among HIV-positive patients in the state of Espírito Santo were not known. In the present study, blood samples from 100 therapy-na?ve HIV-1 infected patients were collected and the HIV subtype was determined through the Heteroduplex Mobility Assay (HMA). Ninety-seven out of 100 studied samples were subtyped by HMA, 73 samples (75.2%) were from subtype B, 9 (9.3%) from subtype F, 3 (3.1%) from subtype C, 6 (6.2%) Benv/Fgag, and another 6 (6.2%) Fenv/Bgag, what suggests that recombinant viruses were present in the studied samples. Twenty-eight percent of the subtype B samples were represented by the Brazilian B" subtype, which were identified by RFLP with Fok I. Data presented here demonstrate that the epidemiological characteristics of the HIV epidemic in the state of Espírito Santo are similar to those from the other Southeastern states and helped to better understand the genetic polymorphism of HIV in Brazil.  相似文献   

13.
白旭华  管永军 《病毒学报》1997,13(4):339-344
应用PCR方法对7份1996年3-5月采集于新疆乌鲁木齐HIV-1阳性静脉吸毒者的外周血单核细胞样品进行扩增,获得了HIV-1膜蛋白基因的核酸片段,并对其C2-V3区及邻区350-450个核苷酸序列进行了测定和分析。  相似文献   

14.
为了解河南郑州市男男同性恋人群HIV-1流行株的亚型分布情况,运用反转录及套式PCR方法从40例已经被确认为HIV-1阳性的MSM的全血样本中扩增gag全长基因并进行序列测定,应用BioEdit软件对序列进行校对编辑,利用MEGA3.1软件构件系统进化树,同时使用美国Los Alamos国家实验室HIV核酸序列库和美国NCBI提供的在线分析工具进行比对分析,确定基因亚型。结果成功获得24条gag基因序列,亚型分析结果显示,共存在B、CRF01-AE和CRF07-BC三种亚型,其中B亚型8例(33.33﹪),CRF01-AE亚型10例(41.67﹪),CRF07-BC亚型6例(25﹪)。小样本量的流行病学调查显示,河南郑州市同性恋人群中主要存在B,CRF01-AE和CRF07-BC三种亚型,CRF01-AE已成为河南郑州市MSM人群中HIV主要流行亚型,河南地区重组型毒株逐渐占优势,流行情况更加复杂。  相似文献   

15.
16.
本研究以一株2006年广东省分离的H9N2亚型禽流感病毒A/Chicken/Guangdong/HL/2006(H9N2)(简称Ck/GD/HL/06)为研究对象,用RT-PCR法扩增病毒基因组各片段(包括5′端和3′端的非编码区序列),将扩增片段进行克隆、测序并与参考毒株的相应序列进行比较分析,绘制各基因片段的系统发生树。分析结果表明,Ck/GD/HL/06株的HA基因同1997年中国香港鸭源毒株Dk/HK/Y280/97(H9N2)在同一进化分支,从HA的糖基化位点、受体结合位点等综合分析,该毒株HA基因未发生明显的变异,符合我国大陆H9亚型禽流感病毒的特点。HA的226位氨基酸残基为亮氨酸(Leu),具有同哺乳动物SAα,2-6受体结合的特性。Ck/GD/HL/06的PB1、PA和NP基因,同2004年越南分离的人源高致病性H5N1亚型流感病毒A/VietNam/1203/2004(H5N1)株(简写A/VN/1203/04)的核苷酸序列一致性分别是93.8%、95%和96.8%,在先前的研究中未见有类似特性毒株的报道,而这种特性H9N2亚型AIV的出现,是否会增加在重组过程中产生新的高致病性H5N1亚型AIV的可能性,是值得我们关注的一个问题,也提醒在我国华南地区应更加重视防控H9N2亚型AIV,做好长期对H9N2亚型AIV监控及分子流行病学调查的工作。  相似文献   

17.
The purpose of this study was to document the genetic diversity of human immunodeficiency virus type 1 (HIV-1) in the Democratic Republic of Congo (DRC; formerly Zaire). A total of 247 HIV-1-positive samples, collected during an epidemiologic survey conducted in 1997 in three regions (Kinshasa [the capital], Bwamanda [in the north], and Mbuyi-Maya [in the south]), were genetically characterized in the env V3-V5 region. All known subtypes were found to cocirculate, and for 6% of the samples the subtype could not be identified. Subtype A is predominant, with prevalences decreasing from north to south (69% in the north, 53% in the capital city, and 46% in the south). Subtype C, D, G, and H prevalences range from 7 to 9%, whereas subtype F, J, K, and CRF01-AE strains represent 2 to 4% of the samples; only one subtype B strain was identified. The highest prevalence (25%) of subtype C was in the south, and CRF01-AE was seen mainly in the north. The high intersubtype variability among the V3-V5 sequences is the most probable reason for the low (45%) efficiency of subtype A-specific PCR and HMA (heteroduplex mobility assay). Eighteen (29%) of 62 samples had discordant subtype designations between env and gag. Sequence analysis of the entire envelope from 13 samples confirmed the high degree of diversity and complexity of HIV-1 strains in the DRC; 9 had a complex recombinant structure in gp160, involving fragments of known and unknown subtypes. Interestingly, the unknown fragments from the different strains did not cluster together. Overall, the high number of HIV-1 subtypes cocirculating, the high intrasubtype diversity, and the high numbers of possible recombinant viruses as well as different unclassified strains are all in agreement with an old and mature epidemic in the DRC, suggesting that this region is the epicenter of HIV-1 group M.  相似文献   

18.
The development of an effective human immunodeficiency virus type 1 (HIV-1) vaccine is likely to depend on knowledge of circulating variants of genes other than the commonly sequenced gag and env genes. In addition, full-genome data are particularly limited for HIV-1 subtype C, currently the most commonly transmitted subtype in India and worldwide. Likewise, little is known about sequence variation of HIV-1 in India, the country facing the largest burden of HIV worldwide. Therefore, the objective of this study was to clone and characterize the complete genome of HIV-1 from seroconverters infected with subtype C variants in India. Cocultured HIV-1 isolates were obtained from six seroincident individuals from Pune, India, and virtually full-length HIV-1 genomes were amplified, cloned, and sequenced from each. Sequence analysis revealed that five of the six genomes were of subtype C, while one was a mosaic of subtypes A and C, with multiple breakpoints in env, nef, and the 3′ long terminal repeat as determined by both maximal χ2 analysis and phylogenetic bootstrapping. Sequences were compared for preservation of known cytotoxic T lymphocyte (CTL) epitopes. Compared with those of the HIV-1LAI sequence, 38% of well-defined CTL epitopes were identical. The proportion of nonconservative substitutions for Env, at 61%, was higher (P < 0.001) than those for Gag (24%), Pol (18%), and Nef (32%). Therefore, characterized CTL epitopes demonstrated substantial differences from subtype B laboratory strains, which were most pronounced in Env. Because these clones were obtained from Indian seroconverters, they are likely to facilitate vaccine-related efforts in India by providing potential antigens for vaccine candidates as well as for assays of vaccine responsiveness.  相似文献   

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