乙、丙型肝炎病毒的分子机制

(一)乙型肝炎病毒基因组由一个不完全的双链 DNA 组成,分子量约1.6×10~6。HBV DNA可分为:①双链松弛状 DNA(RC),有4000个核苷酸;②双链线状 DNA(Linear),有3200个核苷酸;③超螺旋 DNA(SC),含2000个核苷酸;④单链 DNA(SS),含140个核苷酸。其中双链线状 DNA 又分为长链 L(又名负链)和短链 S(又     

与诊断和治疗相关的乙、丙型肝炎病毒研究进展

在引起人类病毒性肝炎的病原中 ,乙型肝炎病毒(HBV)和丙型肝炎病毒 (HCV)由于对人类健康危害较大 ,感染易发展成为肝硬化、肝细胞癌而引起基础和临床研究者的广泛的重视 ,近年来分子生物学研究方法的推广促进了对病毒的研究不断深入 ,从而为进一步发展和完善诊断、治疗提供了新的思路。本文主要介绍近年来一些对诊断和治疗有影响的HBV和HCV病毒学研究进展及对今后发展的可能影响。1　HBV聚合酶基因的变异HBV聚合酶区由A -F 7个基元序列构成 ,催化的核心部位YMDD(酪氨酸 -甲硫氨酸 -天冬氨酸 -天冬氨酸 )四肽基元序列位于C基元序列…     

丙型肝炎病毒及乙型肝炎病毒联合基因免疫实验研究

为ＨＣＶ/ＨＢＶ重叠或混合感染的防治寻求新的高效联合基因免疫策略 ,分别将与ＨＣＶ核心区基因互补的ｃＤＮＡ和ＨＢＶ核心区基因克隆于具有两套独立表达单元的真核表达载体 ｐＲＳＣ的巨细胞病毒启动子和ＲＳＶ启动子下游 ,称为ｐＲＳＣ ＨＢＶ/ＨＣＶ ,转染ＳＰ2 / 0细胞 ,通过免疫荧光和Ｗｅｓｔｅｒｎ印迹法观测蛋白的表达 ,免疫ＢＡＬＢ/ｃ小鼠 ;用酶联免疫法、ＭＴＴ法及荷瘤试验观察小鼠体液免疫应答及细胞免疫应答。结果表明 ,ｐＲＳＣ ＨＢＶ/ＨＣＶ转染ＳＰ2 / 0细胞可见ＨＢｃＡｇ及ＨＣＶ核蛋白染色阳性荧光细胞 ,相对分子质…     

丙型肝炎病毒基因分型的临床应用

丙型肝炎病毒（HCV）感染是输血后肝炎的主要病因之一，50％～70％的急性感染病人将转为慢性肝炎，其中20％的病人将导致肝硬化。目前已知HCV是单股正链RNA病毒，由于HCV复制时所依赖的多聚酶缺乏校对功能，加上为适应环境和逃避宿主免疫监视作用而易于发生突变，形成众多的基因分型。本文仅就有关HCV基因分型与临床应用的研究进展作一综述。一、HCV基因分型现状目前对HCV基因分型的方法多种多样，包括核昔酸序列分析、型特异性引物法、型特异性探针杂交法、限制性片段长度多态性分析和DNA免疫酶学试验等。各种方法在特异性和敏感…     

丙型肝炎病毒基因分型的临床应用

丙型肝炎病毒(HCV)感染是输血后肝炎的主要病因之一,50～100％的急性感染病人将转为慢性肝炎,其中20％的病人将导致肝硬变。目前已知HCV是单     

丙型肝炎病毒基因分型研究

采用逆转录－多聚酶链反应（ＲＴ／ＰＣＲ）法检测了泰安地区４６例输血后丙型肝炎（ＰＴＨ－Ｃ）患者的血清ＨＣＶ－ＲＮＡ〉对其中３４例生进物ＰＣＲ产物用限制性长度多生分析泊（ＲＦＬＰ）进行了ＨＣＶ基因分型。结果ＨＣＶ－Ⅱ型检出率为７３．５％，ＨＣＶ－Ⅲ型检出率为１１．８％；ＨＣＶⅡ／ＨＣＶⅢ型混合感染率为１４．７％，未发现Ⅰ、Ⅳ和Ⅴ型感染。     

甲,乙,丙型肝炎病毒重叠感染

４９３名健康献血员和１９２例ＨＢＶ感染者，用ＥＬＩＳＡ法检测抗－ＨＡＶ－ＩｇＭ和抗一ＨＣＶ，发现献血员之ＨＣＶ感染率为１．６２％。ＨＢＶ感染者的ＨＡＶ、ＨＢＶ二重感染率为１６．７％，ＨＢＶ、ＨＣＶ二重感染率为３．１３％，ＨＡＶ、ＨＢＶ、ＨＣＶ三重感染率为０．５２％。重叠感染者，３２例ＨＡＶ和ＨＢＶ二重感染者，ＨＢｅＡｇ２８例转阴，与１５３例单纯ＨＢＶ感染者５７例转阴比较，有显著性差异（ｐ〈０．０５     

丙型肝炎病毒基因分型的研究进展

丙型肝炎病毒(hepatitis C virus,HCV)属于黄病毒科,分为6个基因型与不同的亚型.HCV的命名系统分为4种,但近年来Simmonds命名系统在国外得到广泛应用.HCV基因分型有明显的地理差异,其流行病学意义较为显著.HCV基因分型的检测方法有5种,每种方法有其各自的优缺点并适合应用于不同的实验研究.HCV基因分型与疾病严重性、病程、病情进展和治疗转归密切相关,但研究者尚未获得一致的意见.不同的HCV基因分型抗病毒治疗的疗效不同,白介素(interleukin,IL)-28B的多态性是不同HCV基因分型治疗后产生持续病毒学应答的预测指标.不同的HCV基因分型给疫苗研制带来一定困难,目前国内外对HCV疫苗的研究在动物身上取得一定成果,现对以上内容作一综述.     

Simultaneous detection of HBV and HCV by multiplex PCR normalization

AIM: To design and establish a method of multiplex PCR normalization for simultaneously detecting HBV and HCV. METHODS: Two pairs of primers with a 20 bp joint sequence were used to amplify the target genes of HBV and HCV by two rounds of amplification. After the two rounds of amplification all the products had the joint sequence. Then the joint sequence was used as primers to finish the last amplification. Finally multiplex PCR was normalized to a single PCR system to eliminate multiplex factor interference. Four kinds of nucleic acid extraction methods were compared and screened. A multiplex PCR normalization method was established and optimized by orthogonal design of 6 key factors. Then twenty serum samples were detected to evaluate the validity and authenticity of this method. RESULTS: The sensitivity, specificity, diagnostic index and efficiency were 83.3%, 70%, 153.3% and 72.2%, respectively for both HBsAg and anti-HCV positive patients, and were 78.6%, 80%, 258.6% and 79.2%, respectively for HBsAg positive patients, and were 75%, 90%, 165% and 83.3%, respectively for anti-HCV positive patients. CONCLUSION: The multiplex PCR normalization method shows a broad prospect in simultaneous amplification of multiple genes of different sources. It is practical, correct and authentic, and can be used to prevent and control HBV and HCV.     

HCV实验室检测方法与临床应用

HCV感染的实验室检测方法包括血清免疫检测和核酸检测。本文对各种方法和不同项目的特点、意义、应用范围及相互关系进行介绍,为临床医生在丙型肝炎诊治时对实验室项目的选择以及结果的解释提供帮助。     

HBV and HCV therapy

One year of interferon therapy inhibits HBV replication in one third of the patients whereas long-term administration of oral nucleos(t)ide analogues is efficient in most of them, as long as early treatment adaptation in patients with partial virological response and resistance is provided. Following the demonstration of a more potent antiviral effect in terms of sustained virological response (SVR) rates, Pegylated-IFN coupled with Ribavirin has become the standard treatment for chronic hepatitis C, with nearly 65% of all treated patients achieving a SVR. Long-term suppression of HBV and eradication of HCV would halt the progression of chronic hepatitis to cirrhosis, hepatocellular carcinoma and liver decompensation.     

应用基因表达谱芯片技术筛选HCV p7蛋白反式调节基因

目的应用基因芯片技术对pcDNA3.1(-)和pcDNA3.1(-)-p7分别转染的HepG2细胞的基因表达谱进行分析,筛选能被HCV p7蛋白反式调节的靶基因,研究p7蛋白的生物学功能。方法以含有HCV全基因组的pBRTM质粒作为模板,应用聚合酶链反应(PCR)扩增p7蛋白编码基因片段,以常规的分子生物学技术构建表达载体pcDNA3.1(-)-p7。以脂质体技术转染肝母细胞瘤细胞系HepG2,提取总mRNA,逆转录为cDNA,与转染空白表达载体pcDNA3.1(-)的HepG2细胞进行DNA芯片分析并比较。结果构建的表达载体经过限制性内切酶分析和DNA序列测定,证实准确无误,提取高质量的总mRNA并逆转录成cDNA,进行DNA芯片技术分析。在1152个基因表达谱的筛选中,有1个基因表达水平显著上调,22个基因表达水平显著下调。结论 HCV p7蛋白是一种反式调节因子,p7基因的表达对于肝细胞基因表达谱有显著影响。     

Post-transplant hepatitis: HCV and HBV

Chronic hepatitis C virus (HCV) is currently the leading indication for liver transplantation (LTx). Hepatitis B virus (HBV) infection occurs less frequently and is associated with better treatment options in the perioperative period. The impact of HCV and HBV in LTx is well recognized. However, therapeutic interventions are less standardized and often depend upon institutional protocol. This article provides a comprehensive review of the literature and addresses many issues and complications with transplantation in patients suffering from chronic liver disease as a result of HCV or HBV.     

Seroepidemiology of HCV and HBV infection in northern China

The HCVAb positive rate in normal population in Beijing was 2.1% and HBsAg positive rate was 2.5%. There is an increasing tendency in the aged group. Plasmapheresis might have been the major cause of HCV transmission in blood donors in the Hebei area. There was a high prevalence of HCVAb and HBsAg in chronic liver diseases in the Beijing area and the HCVAb-positive rate significantly increased corresponding to disease progression.     

HCV相关HCC中伴随HBV感染的意义

目的了解HCV相关HCC中伴随HBV感染的意义,初步探讨HCV和HBV在HCC的发生中的独立作用或相互作用的机制.方法原位杂交检测HCC中c-myc,H-ras,N-ras和nm23H1的mRNA表达,观察它们在伴随HBV感染组(n=44)与单纯HCV感染组(n=10)之间的差异.结果 H-ras和nm23H1在两组的表达强度有差别,H-ras在C+B组的表达强于C组(P=0.0254),而nm23H1的表达C组强于C+B组(P=0.0158).结论 HBV与HCV能协同影响H-ras的表达;单纯HCV相关HCC组之nm23H1 mRNA表达高于伴随HBV感染组,可能是HCV相关HCC低转移率的机制之一.