贵州地区丙型肝炎患者病毒基因型与外周血辅助性 T细胞因子表达相关性研究

目的：了解贵阳地区丙型肝炎患者的HCV基因分型情况,探讨不同 HCV基因分型患者外周血辅助性T细胞因子的差异。方法收集2011年10月至2013年7月在贵阳市第五人民医院住院和门诊的198例慢性丙型肝炎患者样本,分别采用直接测序法检测丙型肝炎基因型,ELISA法检测血清中细胞因子白介素2（IL‐2）、细胞因子白介素8（IL‐8）、细胞因子白介素10（IL‐10）和肿瘤坏死因子α（TFN‐α）的含量。结果198份丙型肝炎患者HCV基因分型结果显示,1a型4例（2．02％）,1b型71例（35．86％）,2a型9例（4．55％）,3a型29例（14．65％）,3b型47例（23．74％）,6a型37例（18．69％）,6d型1例（0．50％）;慢性丙型肝炎患者血清IL‐8与正常对照组比较差异无统计学意义,而慢性丙型肝炎患者血清IL‐2、IL‐10、T N F‐α与正常对照组比较差异有统计学意义;慢性丙型肝炎患者IL‐2与TNF‐α在各基因型间比较差异无统计学意义,lb型患者血清IL‐8与IL‐10含量明显高于3a、3b、6a型患者,差异有统计学意义。结论贵州地区 HCV 基因型分布呈现多样性。贵州地区丙型肝炎1b型患者细胞因子水平失衡可能是导致其慢性化率高的主要因素。     

福建地区慢性丙型肝炎患者丙型肝炎病毒基因分型研究

目的 了解福建地区慢性丙型肝炎病毒(HCV)患者基因型的分布特点.方法 应用反转录聚合酶链反应(RT-PCR)结合荧光探针技术对134例慢性丙型肝炎患者丙型肝炎病毒检测基因分型,并分析基因型与患者年龄、性别以及HCV RNA载量之间的关系.结果 134例血清标本中128例可以分型,检出1a、1b、2a、3a、3b、6a型单基因型(分别为0.7％、45.5％、14.9％、6.7％、2.2％、17.9％)和1b+3b(5.2％)、1b+6a(2.2％)混合基因型共8种,其中以1b型(45.5％)为主,6a型(17.9％)次之.男女慢性丙型肝炎患者均以1型为主要基因型,但差异无统计学意义(x2=0.637,P＞0.05),基因6型男性多于女性,差异有统计学意义(x2=4.666,P＜0.05).基因1型、基因2型平均年龄均高于基因3型、6型、混合型,差异有统计学意义(P＜0.05),而基因1型和基因2型之间,基因3型、基因6型和混合型之间年龄水平相近,差异无统计学意义(P＞0.05).5种基因型HCV RNA载量相近,差异无统计学意义(F=2.157,P＞0.05).结论 福建地区慢性丙型肝炎患者基因分型呈现多样性,1b型流行主导地位呈下降趋势,6a型流行地位处于上升趋势.HCV基因分型在患者性别上分布有所不同,各基因型的患者年龄之间有差别,但各基因型的HCV RNA载量相近.     

辽宁地区丙型肝炎病毒基因分型特点及其临床意义

目的研究辽宁地区丙型肝炎病毒(HCV)基因分型特点,并分析其临床意义。方法选取100例丙型肝炎患者作为样本,首先采用聚合酶链式反应(PCR)-荧光探针法对丙型肝炎患者HCV基因分型进行测定,对PCR法分不出型别的样本,采用测序法检测,对HCV基因分型特点进行观察。结果辽宁地区100例HCV感染患者中,基因型包括1a型、1b型、2a型、3a型、3b型、6a型、6n型7种类型。上述类型中,1b型患者占比最高,1a型患者占比最低。各年龄段中,30~60岁患者各基因型占比更高。1b型、3b型患者,以男性居多;其他基因型患者,以女性居多。1a型患者感染途径以静脉吸毒为主,1b型患者可经血液、静脉吸毒、性传播、母婴传播等多种途径感染,各途径感染风险均较高。2a型患者,感染途径以输血、血制品等院内感染为主。3a型患者感染风险低,感染途径一般为静脉吸毒及性传播。3b型患者,经血液、静脉吸毒感染者居多。6a型及6n型患者,多为血液感染及静脉吸毒感染。结论辽宁地区HCV基因,以1b型基因为主,该基因型可经多种途径感染,临床应以之为重点,对HCV感染进行诊断以及治疗,抑制病情进展,改善患者的生活质量。     

HCVRNA载量在慢性丙肝、肝硬化、肝癌患者中的变化及基因分型

目的了解丙型肝炎病毒RNA（HCVRNA）载量在慢性丙型肝炎、丙型肝炎肝硬化、丙肝相关的肝癌（HCV—HCC）患者中的变化及基因分型对疾病进展的影响。方法回顾性分析2010年2月至2013年8月门诊及住院的1386例HCVRNA阳性且未进行抗病毒治疗的丙型肝炎患者的临床病例资料,把患者按诊断分为丙型肝炎组、丙肝肝硬化组、丙肝相关的肝癌组,采用qRT—PCR检测患者血清的HCVRNA载量与基因分型,多组比较采用方差分析或卡方检验,两组间比较采用t检验。结果丙型肝炎组、丙肝肝硬化组及丙肝相关的肝癌组患者HCVRNA载量分别为（6．47±1．03）lgIU／mL,（6．18±1．09）lgIU／mL和（6．07±1．13）lgIU／mL,三组患者HCVRNA载量不同,丙型肝炎组HCVRNA明显高于其他二种,且差别有统计学意义（F=12．80,P〈0．05）,但丙肝肝硬化组与HCV—HCC组患者HCVRNA载量差别无统计学意义（t=0．65,P〉0．05）。1386例患者中816例进行了HCV基因型检测,1型基因型529例（64．83％）,2型基因型287例（35．17％）,丙型肝炎组患者1型基因型者为400例,2型基因型者为226例,1b型基因型为主（63．10％）,丙肝肝硬化组1型基因型117例,2型基因型为57例,1b型基因型为主（65．52％）,HCV—HCC组1型基因型为12例,2型基因型为4例,1b型基因型为主（68．75％）,三组患者基因型比例无明显差别（x^2=1．02,P=0．31）。结论HCVRNA载量在慢性丙型肝炎发展为肝硬化、肝癌的过程中减低,河南地区丙肝患者以1型基因型为主,且1型、2型基因型对疾病的进展没有影响。     

TaqMan-MGB探针对HCV基因的检测及分型研究

目的建立快速准确检测丙型肝炎病毒（hepatitis C virus,HCV）常见基因分型方法,为丙型肝炎临床诊断和治疗提供依据。方法对40例HCV-RNA阳性血清标本,通过逆转录为cDNA后,进行TaqMan-MGB探针检测、分型。结果 HCV-RNA阳性血清40例,共检出39例,其中1b型29例,占74.36%,2a型5例占12.82%,3b型6例占13.38%。结论所测患者的基因型以1b为主,3b次之,2a相对较低;TaqMan-MGB探针分型的符合率达97.5%。     

吉林地区丙型肝炎病毒基因分型及临床意义的研究

目的了解吉林地区HCV基因分型情况,为HCV感染者的诊断和治疗提供重要的科学依据。方法荧光定量RT-PCR检测165例抗-HCV阳性血清标本,并对153例HCV-RNA阳性的血清标本应用基因芯片法进行HCV基因分型。结果 153例阳性标本中HCV1b型85例,HCV2a型63例,HCV1b/2a混合型5例。结论吉林地区HCV基因型主要为1b型,2a型次之,同时存在HCV1b/2a混合型。     

膜显色DNA芯片技术检测丙型肝炎病毒基因型及临床意义

目的了解丙型肝炎病毒基因型分布及其临床意义。方法应用逆转录-聚合酶链反应(RT-PCR)对89例抗HCV阳性血清进行HCV RNA扩增,再应用膜显色DNA芯片技术,对HCV进行基因分型,并对部分标本用基因测序法作对照。结果89份抗HCV阳性血清中,检测出HCV RNA阳性73例。其中,HCV／1b型66例(90．4％),HCV／1a型1例(1．4％),2a型3例(4．1％),3b型3例(4．1％)。13例标本作基因测序分型对照,分型结果完全一致。15例献血员血清中未检出HCV RNA。结论HCV／1b型是江苏常州地区丙肝病毒优势株。膜显色DNA芯片可用于HCV RNA检测及基因分型检测,方法简便、快速、准确,适合临床推广应用及流行病学研究。     

广东地区丙型肝炎患者HCV基因亚型分析

目的 了解广东地区丙型肝炎患者HCV的基因亚型分布特征,为丙型肝炎的治疗与预防提供依据.方法 采用巢式逆转录-聚合酶链反应扩增广东省194例丙型肝炎患者HCVNS5B基因区域,特异性PCR产物测序后通过美国洛斯拉莫斯国家实验室HCV数据库的Blast程序进行HCV基因亚型分析.结果 194例丙型肝炎患者中有99例为单纯HCV感染者、95例为HIV/HCV共感染者.丙型肝炎患者HCV共有8种基因亚型,其中1b亚型占42.3％(82/194)、6a亚型占35.0％(68/194)、3a亚型占11.9％(23/194)、3b亚型占6.7％(13/194)、2a亚型占2.6％(5/194)、1a亚型占0.5％(1/194)、4a亚型与0.5％(1/194)、5a亚型占0.5％(1/194).丙型肝炎患者中的单纯HCV感染者HCV有7种基因亚型,以1b亚型为主,占67.7％(67/99);丙型肝炎患者中的HIV/HCV共感染者HCV有5种基因亚型,以6a亚型为主,占53.7％(51/95).结论 广东地区丙型肝炎患者HCV基因亚型呈现多样性,主要基因亚型为1b亚型和6a亚型;丙型肝炎患者中的单纯HCV感染者与HIV/HCV共感染者HCV主要基因亚型不同.     

南京地区吸毒人群丙型肝炎的常见基因型分型研究

目的了解南京地区吸毒人群丙型肝炎病毒（hepatitis C virus,HCV）常见基因型分布特点。方法对南京市强制戒毒所收集提供的198份抗-HCV阳性吸毒人员血清进行HCV RNA检测,对阳性标本按照Simmonds分型方法,采用5′非编码区（5′NCR）1、2、3、1b型特异性引物进行PCR扩增与分型,同时分析不同类型吸毒人员丙型肝炎感染者上述基因型分布差异。结果198份血清中,有168份为HCV RNA阳性。单一基因型占77.4%,其中1b亚型106份（63.1%）,2型15份（8.9%）,3型9份（5.4%）;混合基因型占20.2%,其中1b/2型25份（14.8%）,1b/3型5份（3.0%）,2/3型4份（2.4%）;未确定基因型4份（2.4%）。静脉和非静脉吸毒人群HCV基因型分布差异无统计学意义（χ^2=3.614,P〉0.05）,单一基因型和混合基因型分布差异亦无统计学意义（χ^2=0.018,P〉0.05）。结论南京地区吸毒人群HCV基因型1b型为主,其他多种基因型并存,总体分布特点介于中国南北方之间。     

丙肝基因分型研究

目的了解河南地区不同丙型肝炎病毒（HCV）基因型的分布特点,对病情及抗病毒的疗效进行预测。方法应用逆转录-聚合酶链反应（RT—PCR）技术对丙肝患者进行HCVRNA定量检测,对60例HCV—RNA阳性标本应用PCR进行HCV基因分型。采用SPSS10．0进行统计学分析。结果60例HCVRNA阳性的丙肝患者中,1b型46例,占77％,2a型为14例,占23％。结论河南地区HCV感染者中以1b型为主,对HCV感染者在治疗前进行有效的疗效预测具有重要的临床意义。     

我国丙肝病毒结构区的cDNA克隆

目的克隆我国西部丙型肝炎病毒全部结构区基因并进行序列分析,为研究丙肝病毒和基因工程表达做准备。方法从1名西安市丙肝感染者血液中,用Trizol试剂裂解HCV病毒颗粒,用糖原与RNA共沉淀提取HCVRNA,用AMV逆转录酶和随机引物逆转录为cDNA,用RT-PCR方法扩增目的片段并转化到pGEM-T质粒,得到pGEM-T-HCVc,pGEM-T-HCVe1和pGEM-T-HCVe2克隆。将以上3个克隆用特定的酶降解,用连结酶进行连接,构建了HCV结构区的完整克隆。结果将克隆好的质粒从两端双向测序,得到完整的HCV结构基因cDNA核苷酸序列,总长度为2238bp,与已发表的HCV序列长度一致,未发现缺失或移码突变,序列中间亦未发现转录终止子。本文序列与HCV1a,1b序列核苷酸的同源性分别为91.8%,84.5%。氨基酸的同源性分别为94.2%,86.3%。本文序列与HCV1a核苷酸及氨基酸的同源性均大于90%,应为HCV1a亚型。结论我国西部地区存在HCV1a亚型,所克隆HCV结构区cDNA克隆可以用于丙肝病毒研究和基因工程表达。     

Replication priority of hepatitis C virus genotype 2a in a Chinese cohort

HCV genotypes have been documented in clinical practice. The aim of this study was to determine the replication priority of different HCV genotypes in a Chinese HCV positive cohort. Serum samples from 491 apparently healthy Chinese blood donors testing positive for HCV antibodies and naive to antiviral drug therapy were tested. Genotyping analysis showed that genotypes 1b and 2a were predominant and accounted for 77.6% of the HCV infections. Among the genotype groups, individuals infected with genotype 2a had an HCV RNA viral load (10⁸ copies/mL) about 200-fold (lg, 2.3) greater than those infected with other genotypes (10⁴–10⁵ copies/mL) indicating a replication priority of genotype 2a. However, there was no correlation between HCV genotype and antibody response suggesting that the amplification advantage of genotype 2a results from a favorable interaction with the host cellular environment. In conclusion, HCV genotypes 1b and 2a are the predominant genotypes in China and genotype 2a possesses a significant replication priority compared with the other genotypes. This suggests the existence of host cellular factors that may act as drug-targets for entirely clearing HCV infection in the future.Abbreviations: EDTA, ethylenediaminetetraacetic acid; GPT, glutamate-pyruvate transaminase; HCV, hepatitis C virus; NS3, NS4 and NS5, non-structure protein 3, 4 and 5; RdRp, RNA dependent RNA polymerase; SVR, sustained virological response     

粘病毒抵抗蛋白A和HCV基因型与干扰素治疗HCV/HIV患者早期疗效的相关分析

目的 探讨干扰素治疗慢性丙型肝炎合并艾滋病毒感染( HCV/HIV)患者早期疗效与干扰素( INF)诱导的粘病毒抵抗蛋白A(MxA)基因的单核苷酸多态性(SNP)和HCV基因型之间的关系.方法 151例HCV/HIV患者用INFα -2b联合Rib治疗24周,评价早期疗效,根据疗效将其分为早期应答与非应答组.应用PCR及限制片段长度多态性的分析方法,检测患者HCV-RNA基因分型、MxA-88及-123位点的SNP,并分析SNP与INF的早期疗效关系,HCV基因1型与非1型患者MxA基因型之间INF的早期疗效.结果 151例HCV/HIV患者除去12例HCV基因分型无记录者,139例患者中52例为HCV基因1型(34.4％),非1型87例( 57.7％);HCV基因非1型早期应答率(85.1％)高于基因1型(55.8％),差异有统计学意义(x2=14.547,P＜0.001);MxA启动子-88位点,GT型与GG、TT型患者疗效比较,各型间应答率GT型(79.7％)好于GG型(67.7％)及TT型(70.0％),但三者之间差异无统计学意义(x2=2.711,P＞ 0.05).MxA-123位点,CA型、CC型及AA型三者INF疗效应答率分别为78.0％、72.6％及75.0％,差异无统计学意义(x2=0.453,P＞0.05).HCV基因Ⅰ型与非1型患者MxA基因型之间INF的早期应答率比较,HCV基因非1型患者G/T、T/T型(91.5％)好于GG型(77.5％),但差异无统计学意义(x2=3.327,P=0.068),HCV基因1型患者MxA各基因型间应答率差异也无统计学意义.但是,HCV基因非1型患者与Ⅰ型患者间比较,前者G/T、T/T型(91.5％)应答率高于后者G/T、T/T型(62.5％),差异有统计学意义(P＜0.05).结论 HCV基因非1型患者MxA启动子-88、-123各基因型对INF应答率好于HCV基因1型患者,MxA-88位点为GT型的HCV/HIV患者的INF疗效较好,可为INF治疗的预后及个性化治疗提供参考依据.     

Predicting sustained virological response and anaemia in chronic hepatitis C patients treated with peginterferon alfa-2a (40KD) plus ribavirin

AIM: To assess the likelihood of a sustained virological response (SVR) vs. the likelihood of anaemia in patients with chronic hepatitis C. METHODS: Data from 1732 patients treated with peginterferon alfa-2a (40KD) plus ribavirin in two randomized, multinational studies were pooled. Probabilities of SVR and anaemia were modelled using the generalized additive logistic model, with numerous clinical variables considered for entry into the model. Baseline haemoglobin was only considered in the analysis for anaemia. RESULTS: The probability of anaemia increased from 6 to 16% as a function of the ribavirin dose kg(-1) (12-16 mg kg(-1)), whereas the relationship between SVR and ribavirin dose kg(-1) was influenced by hepatitis C virus (HCV) genotype. The probability of an SVR was not influenced by the ribavirin dose kg(-1) in patients with HCV genotype 2 or 3 infection, but increased as a function of ribavirin dose kg(-1) in patients with HCV genotype 1 infection (40-50% increase in probability of SVR for 12-16 mg kg(-1) dose ribavirin increase). The probability of an SVR in patients included with HCV genotype 1 decreased with increasing HCV RNA level to about 3 million copies ml(-1), but was relatively independent of increasing HCV RNA level thereafter. In addition, older age, a higher ribavirin apparent oral clearance and cirrhosis had a negative impact on achieving an SVR, but improved with increasing alanine aminotransferase (ALT) quotient. Sex and ribavirin dose kg(-1) were the most important prognostic factors for anaemia, followed by baseline haemoglobin, age, baseline ALT quotient and cirrhosis. CONCLUSION: This study supports individualizing ribavirin dosages by HCV genotype and body weight, and highlights several clinical variables that influence the likelihood of an SVR compared with anaemia in chronic hepatitis C patients treated with peginterferon alfa-2a (40KD) plus ribavirin.     

重庆地区部分静脉注射毒品者丙型肝炎病毒感染状况的调查

目的··:调查重庆地区部分入住我院的静脉注射毒品者丙型肝炎病毒(HCV)不同基因型感染状况。方法·· :用酶标记免疫(ELISA)法检测HCV感染情况 ,用逆转录 -聚合酶链反应(RT -PCR)与限制性长度多态性(RFLP)分型检测HCVRNA ,并观察血清ALT、AST水平。结果··:入住我院的静脉注射毒品者中抗 -HCVIgG阳性率为40.5 %;HCVRNA阳性率为34.5 %,其中HCV1b型、2a型及1b/2a混合型感染率分别为33.7 %、46.9 %及19.4 %。在ALT/AST正常者和异常者之间 ,HCV各类型感染率无显著性差异。结论··:重庆地区部分静脉注射毒品者HCV感染以2a型为主 ,其次为1b型 ,1b/2a混合型亦不少见。HCV基因型与血清ALT和AST水平变化无明显相关性。     

Implications of rapid virological response in hepatitis C therapy in the US veteran population

Aliment Pharmacol Ther 2012; 35: 105–115

Summary

Background Early predictors of response to hepatitis C virus (HCV) therapy, such as rapid virological response, are valuable for the identification of patients with a higher likelihood of treatment success. Aim To identify predictors of rapid virological response in a real world setting. Methods Using the VA Clinical Case Registry, we identified patients with HCV mono‐infection, without liver transplantation, who initiated peginterferon (PEG‐IFN) and ribavirin (RBV) in 2007 or 2008 and had HCV RNA testing for RVR. Significant baseline characteristics from genotype specific univariate analyses were used in backwards stepwise models to identify significant independent predictors of RVR. Results The final cohort consisted of 2424 patients with genotype 1 (G1), 666 patients with genotype 2 (G2), and 419 patients with genotype 3 (G3). Rapid virological response rates were 15% for G1, 71% for G2 and 57% for G3. Sustained virological response rates were significantly higher in patients with rapid virological response than without, increasing from 18% to 52% in G1, 39% to 71% in G2, and 40% to 60% in G3 (P < 0.0001). A baseline HCV RNA<500 000 IU/mL positively predicted RVR across all genotypes studied. In addition, for G1, Black race, Hispanic ethnicity, aspartate aminotransferase/alanine aminotransferase (AST/ALT) ≥0.6, ferritin≥350 ng/mL, LDL<100 mg/dL and diabetes; for G2, BMI≥30 kg/m², platelets<150 K/μL, LDL<100 mg/dL and the use of PEG‐IFN alfa‐2b; and for G3, AST/ALT≥1.0, all negatively predicted rapid virological response. Conclusion We found several novel independent predictors of rapid virological response, including BMI, AST/ALT ratio, ferritin, platelets, LDL, diabetes and type of PEG‐IFN prescribed, which may be useful in guiding treatment decisions in routine medical practice.     

Systematic review: epidemiology of hepatitis C genotype 6 and its management

Aliment Pharmacol Ther 2011; 34: 286–296

Summary

Background Hepatitis C virus (HCV) genotype 6 is common among patients from Southeast Asia and the surrounding regions, where HCV prevalence is also high. HCV genotype 6 has great genetic diversity and different response to antiviral therapy compared with the more commonly known genotype 1. Aim Our goal was to provide a systematic review of the current literature on the epidemiology, classification and treatment of HCV genotype 6. Methods A search using PubMed for ‘hepatitis C’ AND ‘genotype 6’ produced a total of 47 articles, of which 33 articles were found to be relevant and included in this review. Additional articles were identified using the reference lists of these 33 primary articles. Results The prevalence of HCV genotype 6 is estimated to be as high as 50% in some regions of Southeast Asia with demonstrated significance among intravenous drug users and thalassemia major patients. Although previous line probe assays may have misclassified HCV genotype 6 as genotype 1, newer line probe assays can more accurately and reliably determine HCV genotype. Patients infected with HCV genotype 6 respond better to interferon‐based therapy compared with those infected with genotype 1, although patient baseline clinical characteristics and side effect profiles are similar between HCV genotype 6 and other HCV genotypes. Conclusions Future studies should seek to clarify issues regarding early predictors for treatment response in patients with HCV genotype 6, and the impact of ethnic and genotypic factors to treatment response in HCV genotype 6 patients.