不同人群丙型肝炎病毒基因分型

目的 研究不同人群丙型肝炎病毒的基因分型。方法 对141例抗HCV（ ）、60例抗HCV（-）及28例正常献血员的共132份HCV-RNA阳性血清,运用限制性片段长度多态性分析（RFLP）法作酶切分型。结果 3组人群不同基因型检出率分别是1b/Ⅱ型87.18%、100%、100%;2a/Ⅲ1.71%、0、0;两者混合型（1b/2a型）11.11%、0、0。对抗HCV（ ）血清,分为ALT正常组和异常组,2组人群不同基因型检出率分别是1b/Ⅱ型93.33%、83.33%;2a/Ⅲ型0、2.78%;两者混合型7.14%、13.89%。结论 我国不同人群中HCV感染者均以HCV1b/Ⅱ型为主,今后应以HCV1b/Ⅱ型为重点,加强HCV的诊断、治疗和疫苗研制工作。     

The natural history of hepatitis C virus infection

Chronic hepatitis C virus (HCV) infection is a major health problem worldwide. The natural history of HCV infection is not fully understood. For years, there has been an overestimation of the rate of chronicity in acute HCV. Similar high rates of progression to cirrhosis in chronic HCV were reported. The source of confusion stems from the fact that most acute HCV infections are asymptomatic and never come to medical attention. The consequence of this is that most early studies of natural history reflect the more severe end of the spectrum of the disease. Recent studies reported 43-45% rate of chronicity as opposed to the old rates of 77-85%. Also, the rate of progression to cirrhosis and hepatocellular carcinoma was found to be much lower than previously reported. Multiple factors contribute to the chronicity and progression to cirrhosis, the most important being male gender, age, alcohol intake, and the degree of liver fibrosis on initial biopsy. At least 38% of patients with HCV will manifest symptoms of at least one extrahepatic complication. The most important extrahepatic manifestation is mixed cryoglobulinemia. Other extrahepatic manifestations and their response to antiviral therapy are discussed.     

丙肝病毒核心抗原检测在丙肝病毒感染诊断中的作用研究

目的探讨丙肝核心抗原检测在丙肝感染诊断中的作用。方法收集175例丙肝抗体检测标本同时检测HCVAb、HCV游离抗原、HCV总抗原、HCVRNA。结果 75例HCVAb阳性反应标本,检出HCV游离抗原阳性17例,HCV总抗原阳性54例,HCVRNA阳性50例;100例HCVAb阴性反应标本,检出HCV游离抗原阳性1例,HCV总抗原阳性2例,HCVRNA阳性1例。结论 HCV核心抗原是HCV早期感染的标志之一,检测HCV核心抗原有利于HCV感染的早期诊断。     

丙型肝炎病毒感染的管理

黄病毒属RNA病毒丙型肝炎病毒（HCV）慢性感染是肝脏疾病的一个主要原因。英国卫生部估计，该国HCV慢性感染者约2百万人——其中50％并不知道自身感染了这种病毒。流行情况在不同人群中有所差别：献血者为0．04％，泌尿生殖门诊患者为1％，静脉吸毒者可达到50％。一名全科医师平均处置1800名患者就有8～20名丙型肝炎。如果这些患者得以诊治，一半以上可获病毒清除。我们概括了肝脏病学的这一领域，着重强调HCV感染的危险因素、筛查人群以及慢性感染患者的专业治疗。     

Detection of hepatitis G virus RNA in hepatitis C patients and hepatitis C virus infected hemodialysis patients

AlthoughsensitiveandspecificimmunoassayandmolecularbiologicaltechniquesforthedetectionofthehepatitisA--Evirusesareavailable,theetiologyofasubstantialfractionofpost--transfusionandcommunity--acquiredhepatitiscasesremainsundefined[1],suggestingtheexistenceofadditionalcausativeagents.Anewhumanhepatitisviruswasisolatedbytwoindependentgroups.ThenewvirusisprovisionallydesignatedashepatitisGvirus(HGV)[ZJorGBvirusC(GBV~C)[3'4),whichwasthoughttobetheagentofpartofthenon--A--Ehepatitispatients.Fro…     

丙型肝炎病毒C区的DNA改组

目的：利用DNA改组技术进行不同基因型别丙型肝炎病毒（HCV）基因组C区的人工进化。方法：首先利用PCR扩增了三段具有较高序列同源性的460bp基因片段，然后将其等量混合，在Mg^2＋存在的条件下，用脱氧核糖核酸酶（DNase Ⅰ）切割成50bp左右的小片段。这些小片段在不外加引物的条件下，利用PCR反应进行重聚，再将重聚物经过一轮正常的PCR扩增。结果：获得了与原片段大小相当的基因片段。结论：这一技术为进一步筛选高活性的HCVC区基因打下基础，有利于从一组序列同源性程度较高的基因库构建随机嵌合基因，并为改组其他基因家族提供了借鉴。     

DC-SIGN: binding receptors for hepatitis C virus

Objective To review the recent developments in and research into binding receptors of hepatitis C virus (HCV) and especially the role of dendritic cell-specitic adhesion receptor (DC-SIGN) in HCV. Data sources Both Chinese- and English-languge literature was searched using MEDLINE (2000-2003) and the databank of Chinese-language literature (2000-2003). Study selection Relevant articles on DC-SIGN and HCV binding receptors in recent domestic and foreign literature were selected.Data extraction Data were mainly extracted from 40 articles which are listed in the references section of this review. Results DC-SIGN, a dendritic cell-specific adhesion receptor and a type Ⅱ transmembrane mannose-binding C-type lectin, is very important in the function of dendritic cells (DC), both in mediating naive T cell interactions through ICAM-3 and as a rolling receptor that mediates the DC-specific ICAM-2-dependent migration processes. It can be used by HCV and other viral and bacterial pathogens including human immunodeficiency virus (HIV), Ebola virus, CMV and Mycobacterium tuberculosis to facilitate infection. Both DC-SIGN and DC-SIGNR can act either in cis, by concentrating virus on target cells, or in trans, by transmission of bound virus to a target cell expressing appropriate entry receptors. Recent report showed that DC-SIGN not only plays a role in entry into DC, HCV E2 interaction with DC-SIGN might also be detrimental to the interaction of DC with T cells during antigen presentation. Conclusions DC-SIGNs are high-affinity binding receptors for HCV. The clinical strategies that target DC-SIGN may be successful in restricting HCV dissemination and pathogenesis as well as directing the migration of DCs to manipulate appropriate immune responses in autoimmunity and tumorigenic situations.     

瘐型肝炎病毒的分子生物学研究

目的：了解瘐型肝炎病毒的国内外研究历史及在分子生物学特征和实验室诊断方面的最新研究进展。方法：以瘐型肝炎病毒（ＧＢｖｉｒｕｓ Ｃ／ｈｅｐａｔｉｔｉｓ Ｇ ｖｉｒｕｓ）作为检索词，通过微机，在中国生物医学文献光盘数据库和ＭＥＤＬＩＮＥ光盘数据库中检索９５年以后庚型肝炎病毒研究方面的有关文献，并查阅原文，认真综合分析，结果：庚型肝病毒是１９９５年发现的一种新病毒，因其可能与人类肝炎相关，而引起生物医学界的广泛关注，目前，国内外已生并测定了不同庚型肝炎病毒株的全基因组序列，陆续开展了其分子生物学特征和实验室诊断方面的研究，并初步建立了检测ＨＧＶ感染的ＲＴ－ＰＣＲ法及酶联免疫法，结论：庚型肝炎病毒的有些基因结构与功能，基因变异与分型还不能完全定论，ＲＴ－ＰＣＲ检测方法和酶联免疫法的灵敏度或特异度还相对较低，尚需进一步研     

Human immunodeficiency virus and hepatitis C virus co-infection

Hepatitis C virus (HCV) infection is a major cause of liver disease and hepatocellular carcinoma worldwide, as well as the leading cause of liver transplantations in the United States. As a result of similar modes of transmission, approximately 30% of HIV-infected individuals are co-infected with HCV. Among intravenous drug users, almost 90% of people infected with HIV are also infected with HCV. Because of treatment with highly active anti-retroviral therapy, HIV-infected individuals have improved survival and are no longer suffering from opportunistic infections and malignancy as in years past. As a result, co-infection with HCV has now become a frequent cause of morbidity and mortality in HIV-infected individuals. Furthermore, liver disease secondary to HCV infection is now the leading cause of hospital deaths in HIV-infected people in the US. HIV infection accelerates the course of HCV-related liver disease and viremia. It is less clear whether HCV infection affects the clinical course of HIV; however, HCV-related liver disease can limit many individuals from receiving anti-HIV therapy. HIV/ HCV co-infection is common, and serious. Physicians caring for HIV-infected patients worldwide must now address hepatitis C virus co-infection.     

丙型肝炎病毒母婴传播的研究

目的:研究丙型肝炎病毒(HCV)母婴传播机率及感染方式.方法:用ELISA和RT-PCR法检测母婴血清及母亲乳汁、羊水、唾液的抗-HCV IgG及HCV-RNA,循环测序法测定母婴HCV-cDNA序列.结果:对1 521例孕妇筛查,其中15例抗-HCV IgG阳性(12例HCV-RNA阳性).所生15例婴儿,随访结果仅1例抗-HCV IgG持续阳性,出生时2例血清HCV-RNA阳性者中的1例抗-HCV IgG及HCV-RNA持续阳性.HCV母婴传播率为16.7%(2/12).其中一对母婴间HCV-cDNA序列同源性为100%.羊水、乳汁、唾液抗-HCV及HCV-RNA检出率分别为100%和25.0%,53.5%和16.7%,100%和0.0%.结论:HCV母婴传播可能发生于宫内或分娩时,羊水可能是引起HCV母婴传播的重要因素,乳汁、唾液引起传播可能性较小.     

加强对乙型肝炎和丙型肝炎病毒准种的研究

生物的变异是生物为了适应环境的不断变化，维持生存和种族繁衍的需要而采取的一种十分重要和必要的应对措施，也是生物进化的基础，这是生物界普遍存在的现象。     

广州地区丙型肝炎患者HCV基因分型研究

目的研究广州地区丙型肝炎患者HCV基因型分布的变化情况。方法收集207例HCV RAN阳性丙型肝炎患者血清,对其HCV Core区(C区)和NS5B区基因序列进行RT-nPCR扩增,两区扩增成功的PCR产物进行基因测序,测序结果分别与相应的参照株序列比对,构建系统进化树,两区基因分型结果一致者确定HCV基因亚型。结果 185例HCV C区和NS5B区都扩增成功,其中179例两区基因分型结果一致。共检出1、2、3、6型4种基因型和8种基因亚型,其中1b型占40.78%(73/179),6a型占27.37%(49/179),3b型占11.73%(21/179),3a型占8.94%(16/179),2a型占6.15%(11/179),1a型占3.91%(7/179),2b型占0.56%(1/179),6n型占0.56%(1/179)。1b株可分为A、B、C三大簇,A簇与中国广泛流行1b株接近、B簇与中国中部及南部地区流行1b株接近,6a株分为Ⅰ、Ⅱ两大簇,与献血人群及静脉吸毒人群中6a型株接近。结论广州地区丙型肝炎患者HCV基因型以1b、6a、3型为主,存在多种亚型;1b型株包含中国大部分地区的1b型株,6a、3型已取代2a型分别成为第二、三主要流行基因型;未发现6型变异株。     

EB病毒转化的丙型肝炎病人外周血B淋巴细胞中丙型肝炎Ⅱ型？…

目的 观察丙型肝炎病人外周血Ｂ细胞长期存活状态下其中是否仍有丙型肝炎病毒（ＨＣＶ）存在。方法 用ＥＢ病毒感染并使ＨＣＶ阳性外周血Ｂ细胞转化,获得含ＨＣＶ的外周血永生化Ｂ细胞（ＬＣＬ）。而后,每月应用套式ＲＴ－ＰＣＲ检测一次培养细胞和上清中的ＨＣＶ－ＲＮＡ,并用酶切法进行ＨＣＶ基因分型,电镜和免疫电镜观察ＨＣＶ在该细胞中存在的部位及其形态结构特征。结果 经连续１年检测,ＨＣＶ在传代细胞中持续存在,培     

丙型肝炎患者血清中丙型肝炎病毒核酸含量与丙型肝炎病毒抗体和丙氨酸氨基转移酶的相关性

[目的]探讨丙型肝炎患者血清中丙型肝炎病毒核酸(HCV RNA)含量与丙型肝炎病毒抗体(抗-HCV)及丙氨酸氨基转移酶(ALT)间的相关性.[方法]采用实时荧光定量PCR检测302例丙型肝炎患者血清中HCV RNA含量,同时采用ELISA法检测抗-HCV水平,利用全自动生物化学检测仪测定ALT水平.[结果]302例丙型肝炎患者血清标本中,抗-HCV均呈阳性,其中199份标本HCV RNA含量高于检测上限(103 U/mL),2种检测方法的符合率为65.9%.ALT异常率随HCV RNA含量的增高而升高,两者呈显著正相关(r=0.155,P<0.05);而ALT水平变化与HCV RNA含量无相关性(r=0.001,P>0.05).[结论]HCV RNA含量与抗-HCV水平是诊断HCV感染的重要指标,HCV RNA含量结合ALT水平测定可帮助临床了解HCV在体内复制的状况及肝脏损伤情况.     

丙型肝炎病毒感染细胞模型的研究概况

丙型肝炎病毒(HCV)是输血后肝炎的重要病原体,近年来对HCV的研究日益受到重视.但由于HCV是一种低复制病毒,实验动物及患者血清和组织中含量较少,直接研究HCV复制表达的机制十分困难,因此迫切需要建立HCV的细胞模型.     

丙型肝炎患者血清中丙型肝炎病毒交叉中和抗体的检测

[摘要]目的分析HCV感染者血清中是否存在交叉反应性中和抗体。 方法以分泌表达HCV包膜E2蛋白真核表达质粒转染的293T 细胞培养上清液中的HCV E2蛋白作为检测抗原,建立检测HCV E2抗体的ELISA方法,检测32份HCV抗体阳性的慢性丙肝患者血清,然后用免疫荧光分析血清与HCV全长包膜蛋白表达质粒转染的293T细胞的结合反应,再用5株HCV假病毒(HCVpp)及两株细胞培养产生的HCV(HCVcc)为模型分析血清的病毒中和活性。 结果32份HCV抗体阳性血清中,26份血清可检测出E2抗体,阳性率81.3%。其中HCV RNA阳性的12份血清均为E2抗体阳性,E2抗体水平与HCV RNA水平负相关。HCV E2抗体阳性血清对5株HCVpp以及两株HCVcc的感染性均有不同程度的中和作用,中和活性与E2抗体水平相平行。结论HCV感染可诱导保护性体液免疫应答,丙肝患者血清中存在交叉中和抗体,提示开发能诱导广泛交叉中和抗体的丙肝疫苗具有可行性。     

肝炎基因诊断芯片对丙型肝炎病毒基因分型检测的初步研究

目的 研究丙型肝炎病毒（HCV）基因分型诊断芯片的制备和其对丙型肝炎患者血清HCVRNA基因分型诊断的准确性。方法 根据丙型肝炎病毒5′端非编码区末端（5UTR）和C区设计引物和特异性探针,将设计的20条特异性寡核苷酸探针用核苷酸合成仪合成后,配制成浓度为50μmol/ml的点样液,用点样仪点到特殊处理的玻片介质上,制成丙型肝炎病毒基因分型诊断芯片,收集HCVRNA阳性的丙型肝炎患者血清60份作为实验组,健康人血清60份作为对照组,用荧光定量PCR仪对两组血清进行HCVRNA定量检测,实验组血清HCVRNA含量均大于500拷贝/ml,对照组血清HCVRNA含量均小地500拷贝/ml,两组血清进行HCVRNA分离纯化,逆转录反应,巢式PCR扩增后,分别用基因芯片,HCVRNA核酸序列测定法（美国Li-Cor核酸序列测定仪）进行双盲丙型肝炎基因分型检测。结果 实验组血清基因诊断芯片检测均为阳性;基因分型;1b型46例,3a型3例。3b型3例,2a型2例,2b型2例,混合型1b 2a2例。1b 3b型2例;核酸序列测定分型1b型50例,3a型3例,3b型3例,2a型2例,2b型2例;基因诊断芯片检测和核酸序列测定检测的基因分型结果符合率为93.3%。对照组血清基因芯片检测匀阴性。结论 肝炎病毒基因诊断芯片可以用于检测血清HCVRNA,并进行基因诊断分型,准确率较率。