慢性丙型肝炎患者病毒基因分型及HCV RNA相关性研究

目的：探讨江苏地区慢性丙型肝炎患者病毒基因分型特点及其与患者年龄、性别和丙型肝炎病毒（HCV）RNA水平的关系。方法收集抗 HCV 阳性血清1032份,采用荧光定量 PCR（Q‐PCR）法检测患者血清HCV RNA水平,HCV基因分型芯片进行基因分型检测,应用SPSS19．0软件对数据进行统计分析。结果检测出HCV RNA阳性样本704份,其中 HCV 单基因型674份,1b、2a、3b、3a、1a 和6型分别占75．71％、7．95％、7．95％、1．99％、0．99％和1．14％;混合基因型30份,1b／2a、1b／3b、1b／3a和2a／3b型分别占2．27％、1．56％、0．28％和0．14％。不同基因型均以男性和青壮年居多,且病毒载量差异有统计学意义：1b和3a相近,都高于2a和1b／2a病毒载量。结论江苏地区 HCV基因型以1b型为主,其次是2a、3b型。慢性丙型肝炎患者以中青年男性居多,而且不同 HCV基因型间病毒载量有差异。     

国产HCV RNA分型试剂盒检测结果分析

目的分析丙型肝炎病毒(HCV)RNA分型试剂盒检测深圳市抗-HCV阳性献血者结果。方法收集2014-2015年158份深圳市抗-HCV阳性献血者血液样本,应用聚合酶链反应(PCR)-荧光探针法进行HCV RNA定量检测,病毒载量1.0×103 IU/mL的样本经HCV RNA分型试剂盒检测HCV基因型,分析不同基因型所占比例,病毒基因型与载量之间的相关性。结果 158份抗-HCV阳性献血者PCR-荧光探针法检出HCV RNA阳性样本54例,病毒载量1.0×103 IU/mL的45例,全部得到分型结果,HCV 1b型、2型、3型、6型分别占57.78%(26/45)、6.67%(3/45)、8.89%(4/45)、26.67%(12/45)。单因素方差分析结果显示,1b型与2型病毒载量差异有统计学意义(F=2.861,P0.05);不同性别间HCV RNA定量检测结果及抗-HCV S/CO值结果差异有统计学意义(P0.05);不同年龄段各基因型分布比例经Fisher精确检验,差异有统计学意义(P0.05)。结论 HCV 1b型、6型仍为深圳市无偿献血人群感染HCV的主要基因型,而HCV 2型和3型比例有所减少。     

某地区丙型病毒性肝炎患者HCV基因分型研究

目的 了解该地区丙型病毒性肝炎(以下简称丙型肝炎)患者丙型肝炎病毒(HCV)基因分型情况.方法 分别以ELISA和重组免疫印迹试进行血清标本抗HCV抗体筛查和确认,对抗HCV抗体阳性标本用实时荧光定量PCR进行病毒载量检测.对病毒载量大于103 copy/mL的标本用多重PCR进行HCV基因分型.结果 125例抗HCV抗体阳性标本中,HCV基因型主要为1b、2a、3a、1c、2c的检出率分别为58.4%、21.6%、3.2%、4.0%和8.0%.结论 该地区HCV感染主要以1b型为主,其次是2a和3a型,检出其他地区少见的1c和2c型,说明该地区丙型肝炎流行的基因型呈多样性.     

新疆地区慢性丙型肝炎患者病毒基因型、病毒载量与HCV抗体S/CO值、肝酶、外周血T淋巴细胞亚群的相关性分析

目的探讨新疆地区慢性丙型肝炎患者病毒基因型、病毒载量对丙型肝炎病毒(HCV)抗体S/CO值、肝酶、外周血T淋巴细胞亚群的影响。方法收集2014年1月至2015年9月新疆医科大学第一附属医院慢性丙型肝炎住院患者106例、体检正常者(正常对照组)30名,分别采用化学发光法、酶法、荧光定量逆转录聚合酶链反应(PCR)、PCR-反向点杂交技术和流式细胞术对106例慢性丙型肝炎患者的血清样本进行HCV抗体、肝酶[丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)]、HCV RNA、HCV基因分型以及外周血T淋巴细胞检测。结果 1b、2a、3a和3b基因型慢性丙型肝炎患者间HCV抗体S/CO值、ALT、CD8+T淋巴细胞百分数差异有统计学意义(P0.05)。病例数较多的1b、2a基因型慢性丙型肝炎患者CD4+T淋巴细胞百分数、CD4/CD8比值均低于正常对照组[CD4+T淋巴细胞百分数分别为(39.05±9.42)%、(38.65±10.26)%、(42.45±4.61)%,CD4/CD8比值分别为1.60±0.64、1.53±0.66、2.31±0.25],CD8+T淋巴细胞百分数[(26.79±7.58)%、(27.15±10.40)%]则高于正常对照组[(18.36±3.88)%,P0.05]。将慢性丙型肝炎患者按HCV RNA载量水平分为4组,各组间HCV抗体S/CO值、肝酶水平及CD4/CD8比值差异有统计学意义(P0.05)。结论新疆地区慢性丙型肝炎患者病毒基因型以1b、2a型为主,不同基因型及病毒载量患者HCV抗体S/CO值、外周血T淋巴细胞亚群及肝酶水平存在差异,具有一定的相关性。     

山东地区448例丙型肝炎患者HCV基因分型状况分析

目的研究丙型肝炎病毒(HCV)基因分型,了解山东地区丙型肝炎的基因分型情况。方法收集448例丙型肝炎患者的血清标本,采用焦磷酸测序法对HCV感染标本进行分型。结果 448例丙型肝炎患者共检出7种基因亚型,分别为1b型288例(64.28%)、2a型137例(30.57%)、1a型13例(2.90%)、3b型4例(0.90%)、3a型2例(0.45%)、6a型2例(0.45%)、1a1b型2例(0.45%);对HCV基因型与性别、年龄的关系进行分析显示无明显相关性;2013-2016年各基因型患病率分析结果显示无明显的变化趋势。结论山东地区HCV感染基因型以1b型为主,其次为2a型;HCV基因型分布与性别、年龄差异无统计学意义(P0.05),且近4年山东地区各基因型年患病率无明显变化趋势。     

肝脂肪变性促进慢性丙型肝炎病程进展的研究

目的 研究脂肪变性在慢性丙型肝炎疾病进程中的作用.方法 收集治疗前行肝穿刺病理检查的慢性丙型肝炎患者159例,按照感染HCV基因亚型分为基因1b型组、基因2a型组和其他基因型组,荧光定量PCR法检测所有病例血清HCV载量,组织学评估各组穿刺肝组织炎症坏死、纤维化和肝细胞脂肪变性程度.结果 HCV基因1b型和2a型感染患者占总病例的65.41%,63.52%(101/159)的慢性丙型肝炎患者发生肝细胞脂肪变性,不同基因型组间脂肪变性程度无统计学差异.不同基因型组HCV慢性感染者间炎症活动度、纤维化程度和脂肪变性程度无统计学差异(P均>0.05).肝脂肪变性与肝纤维化和炎症活动度均密切相关(r值分别为0.34和0.29,P均<0.01),但HCV感染病毒量与脂肪变性、肝纤维化和炎症活动度间无明显相关性.结论 脂肪变性促进慢性丙型肝炎病程进展.     

丙型病毒性肝炎病毒基因型与病毒载量及疾病进展的相关性分析

目的 分析丙型病毒性肝炎(丙肝)病毒(HCV)的基因型与病毒载量及肝脏疾病进展的相关性. 方法 选择2010年12月至2013年4月在宁波市第二医院北郊院区就诊的慢性HCV感染者,分别采用反转录-聚合酶链反应(RT-PCR)法和基因芯片法检测HCV RNA和HCV基因型. 结果 107例标本中,共检出7种亚型,其中1a型4例(3.74%),1b型52例(48.60%),2a型15例(14.02%),3a型10例(9.35%),3b型9例(8.41%),6型16例(14.95%),1b+2a混合型1例(0.93%);男女慢性HCV感染均以1型为主要基因型,但男性6型的构成比高于女性(2=4.336,P=0.049);各年龄组间HCV基因型的构成及各基因型的年龄构成差异均无统计学意义;6型的病毒载量最高,其次为1型,2、3型的病毒载量明显低于6型和1型(P0.01);终末期肝病患者和伴存免疫缺陷患者的病毒载量均高于慢性丙肝患者,且终末期肝病患者以基因1型(主要为1b)为主(P=0.016). 结论 本地区HCV基因型同样以1b型为主,且1b型感染者的病毒载量较高,疾病进展较快;6型的感染率较高,应重视对基因6型HCV感染者的研究.     

茂名地区无偿献血者72例HCV感染特征及检测效果评价分析

目的测定茂名地区部分丙型肝炎病毒感染献血者中不同感染状态比率及其分子特征,分析丙型肝炎病毒感染基因型分布流行特征及趋势,评价ELISA法检测血液HCV的效果。方法对本地区献血者中抗-HCV检测呈反应性的血液标本采用定量PCR方法检测血浆中病毒载量,同时采用巢式PCR技术扩增核酸片段,进而将血液标本分为病毒自然清除(RNA-/抗-+)与病毒血症(RNA+/Ab抗-+)2种状态,并分析2种状态在献血者性别方面的差异。HCV RNA阳性标本通过分析5’-NCR序列进行基因分型。比较ELISA与NAT检测方法的符合性。结果72份抗-HCV呈反应性的标本中,病毒自然清除50份(69.4%),病毒血症22份(30.6%)。22份PCR定量阳性标本中基因分型阳性17份,其中基因1型占17.6%(3/17),基因2型占11.8%(2/17),基因3型17.6%(3/17),基因6型47.1%(8/17),无法准确定型5.8%(1/17)。试剂A检测呈反应性标本中核酸确认阳性标本占32.3%;试剂B为54.5%。女性病毒血症的比例(2/19)比男性(20/53)低27.2%(χ2=9.637,P0.05)。结论茂名地区献血者HCV感染病毒自然清除率约为69.4%,HCV基因型中6型为主要基因型,女性献血者更容易清除病毒。现行2遍ELISA检测策略安全有效,但同时存在特异性不高的弊端,实施ELISA加NAT检测的策略,更能保证血液的安全和降低检测假阳性。     

NS5B区核酸序列测定的丙型肝炎病毒基因分型研究

目的了解昆明市丙型肝炎感染者HCV基因型分布特征。方法用RT-PCR法扩增40例HCVRNA阳性的患者血清样本丙型肝炎病毒NS5B区段,PCR产物纯化后进行测序分析,测序结果与参考序列共同构建系统进化树,得到丙型肝炎病毒基因型和基因亚型。结果丙型肝炎病毒NS5B区段基因序列的系统进化树分析能对丙型肝炎病毒感染者样本进行很好的分型;分型结果显示,昆明地区丙型肝炎病毒主要的基因型是3b型占45.0%,其次2a型占22.5%,1b型占20.0%,3a型占7.5%,6n型及6a型各占2.5%。结论构建丙型肝炎病毒NS5B区系统进化树分析能得到准确的HCV基因型和亚型;昆明地区的丙型肝炎病毒基因型以3型为主,其次2a、1b型,同时发现6型;昆明丙型肝炎感染者中HCV基因型分布更接近于中国香港和东南亚国家的丙型肝炎病毒基因分布。     

甘肃地区藏族丙型肝炎患者的HCV基因分型研究

目的 了解甘肃地区藏族丙型肝炎患者的HCV基因分型特征.方法 对HCV感染的患者样本用实时荧光定量PCR进行病毒载量的确定.对病毒载量大于103 copy/mL的样本用多重PCR进行HCV基因分型.结果 通过对甘肃地区藏族HCV感染者的样本进行基因分型,发现该民族HCV感染以1b型(56.6%)为主,HCV感染2a型(23.3%)次之,再次为2c型(10.0%)和1c型(3.3%),未检测到3a型.结论 甘肃藏族HCV感染以1b型为主,该研究为临床针对不同基因型HCV感染者的治疗提供了依据.     

Estimation of two real-time RT-PCR assays for quantitation of hepatitis C virus RNA during PEG-IFN plus ribavirin therapy by HCV genotypes and IL28B genotype

Hepatitis C virus (HCV) RNA values measured with two real-time PCR methods (Cobas Ampliprep/Cobas TaqMan, CAP/CTM, and the Abbott real-time PCR test, ART) vary among patients with genotype 1. We investigated HCV RNA values measured by two real-time PCR assays during pegylated interferon plus ribavirin (PEG-IFN/RBV) therapy. We evaluated 185 cases of chronic hepatitis C patients, among which 97 patients received the PEG-IFN/RBV therapy. HCV RNA values of CAP/CTM for genotype 1 were significantly higher than those of ART (p < 0.05) The difference in HCV RNA values (CAP/CTM minus ART) of genotype 1 was significantly higher than those in genotype 2 (p < 0.0001). The positive rate (>0) of the difference of HCV RNA values in genotype 1 was 100 % (55/55), which was significantly higher than the 78.6 % (33/42) of genotype 2 (p < 0.001). There was no difference between TT and TG/GG genotype groups in terms of difference of HCV RNA values (CAP/CTM minus ART). After PEG-IFN/RBV therapy was administered, reduction of HCV measurements was observed from day 1 for both assays regardless of genotype. The HCV value of CAP/CTM during PEG-IFN/RBV therapy was consistently higher than the value of ART, although the difference in these two values gradually became smaller during the course of therapy, and eventually no significant difference was observed near the detection level. No correlation was observed between the sustained virological response (SVR) rate and the difference between the CAP/CTM HCV values and the ART HCV value before treatment.     

浙江省湖州地区丙型病毒性肝炎病毒基因型分布

目的 了解浙江省湖州地区丙型病毒性肝炎(丙肝)病毒 (hepatitis C virus,HCV) 基因型的分布特点。 方法 应用反转录-巢式聚合酶链反应(RT-nest PCR)产物直接测序方法对湖州地区94例丙肝病毒核糖核酸(RNA)阳性患者血清丙肝病毒进行基因分型,同时检测其HCV RNA含量和谷丙转氨酶(ALT)水平。 结果 94例患者中,HCV 1a型5例 (5.3%);1b型例77例(81.9%);2a型9例(9.6%);3a型1例(1.1%),6a型2例(2.1%)。各基因型在男女性分布上差异无统计学意义(2＝1.721,P=0.787);1b型和非1b型平均年龄水平差异无统计学意义(t=0.453,P=0.652);血源性感染和非血源性感染患者的基因型分布差异也无统计学意义(2＝0.851,P=0.931),1b型和2a型及其他基因型患者在HCV RNA和ALT水平差异无统计学意义(F=1.078,P=0.345; F=0.867,P=0.424)。 结论 湖州地区HCV基因型主要为1b型,其次为2a型,同时还存在1a型、3a型和6a型等基因型。     

Response to combined interferon and ribavirin is better in patients infected with hepatitis C virus genotype 6 than genotype 1 in Hong Kong

Data on the treatment efficacy of interferon (IFN)-alpha and ribavirin in patients with chronic hepatitis infected with hepatitis C virus (HCV) of genotype 6 are lacking. A study has been reported from Hong Kong which compared the treatment efficacy of IFN-alpha and ribavirin in the treatment of chronic HCV infection with genotypes 1 and 6. Twenty-four patients with HCV genotype 1 and 16 patients with HCV genotype 6 were studied. The baseline demographic data including median age, gender ratio, alanine aminotransferase levels, bilirubin levels, HCV RNA levels and histological scores were comparable between the two groups of patients. All patients received IFN-alpha 5 million units three times per week and ribavirin (1,000 mg for those weighing 75 kg) for 1 year. Patients infected with HCV genotype 6 achieved virological response significantly higher than those with HCV genotype 1 (67 vs. 33% at week 24, p = 0.02; 75 vs. 42% at the end of treatment, p = 0.05; 63 vs. 29% at 6 months after completion of treatment, p = 0.04). Histological improvement in inflammatory activity and fibrosis in the liver were observed in 25% and 25% of patients infected with HCV genotype 6 in contrast to only 13 and 8% in patients infected with HCV genotype 1; however, the differences were not statistically significant. In conclusion, patients with HCV genotype 6 gain a better response to combined treatment with IFN-alpha and ribavirin than those with HCV genotype 1 and achieve a significantly higher rate of sustained virological response.     

丙型肝炎病毒反向斑点杂交基因分型检测

目的:分析厦门地区及肝病患者不同临床类型丙型肝炎病毒(HCV)感染株的基因型.方法:用反向点杂交技术(RDB)对109例HCV RNA阳性的血清进行基因分型.结果:109例HCV RNA阳性血清中,其中单一型感染者98例(89.91%),混合型感染11例(10.09%),单一型感染者中以1b型占绝对优势,共90例(82.57%),2a/2c及6a型分别为2例和3例,另有3例1b+2a/2c和8例1b+6a混合型;HCV 1b型在急、慢性肝炎、肝硬化及肝癌患者中均占绝对优势,其余有少量2a/2c、6a及某些混合感染型.结论:厦门地区HCV感染者中,以HCV 1b基因型为主,其次为HCV 6a,另有少数2a/2c型.不同基因型在不同临床类型中的分布无明显差异.     

Clinical performance of the new rRoche COBAS TaqMan HCV Test and High Pure System for extraction, detection and quantitation of HCV RNA in plasma and serum

We evaluated the Roche COBAS TaqMan HCV Test For Use With The High Pure System (TaqMan HPS; Roche Diagnostics), for the extraction, detection and quantitation of hepatitis C virus (HCV) RNA in serum or plasma of HCV-infected individuals. The TaqMan HPS is a real-time PCR assay with a reported linear dynamic range of 3.0x10(1) to 2.0x10(8) HCV RNA IU/ml, and a reported lower limit of detection (LLD) of 10 IU/ml. Calculation of the HCV RNA titre is based upon an external standard curve in the presence of an internal control. Intra-assay and inter-assay variation were small in reference panel members with HCV RNA > or =100 IU/ml. Genotype performance and quantitative correlation between the TaqMan HPS and the bDNA (VERSANT HCV 3.0 assay; Bayer Diagnostics), assessed in 59 patient samples, were good for HCV genotype 1 but poor for genotypes 2, 3 and 4. For genotypes 2, 3 and 4, values obtained from the TaqMan HPS were in general 0.5 log lower than those from the bDNA. Sensitivity was poor in low viral titre samples of genotypes 1, 2, 3 and 4. The LLD (95%) was estimated at 41 HCV RNA IU/ml for genotype 4. The TaqMan HPS underestimates HCV RNA at all levels in plasma and serum from HCV-infected individuals, and the LLD should be reconsidered. This is clinically relevant because underestimation of HCV RNA levels during therapy may lead physicians into making incorrect treatment decisions.     

Mixed cryoglobulinaemia in patients with chronic hepatitis C infection: prevalence, significance and relationship with different viral genotypes

In order to analyse the prevalence and significance of cryoglobulinaemia in patients with chronic hepatitis C virus (HCV) infection and the possible relationship of cryoglobulinaemia with the genotypes of HCV, we studied 89 patients with chronic HCV infection, 42 healthy controls and 22 patients with alcoholic cirrhosis. The patients with HCV were divided into three different groups according to the presence of cirrhosis and alanine aminotransferase levels. Moreover, in 20 patients with HCV and cryoglobulinaemia, HCV RNA sequences were quantified in serum and in cryoprecipitate. Cryoglobulins were detected more frequently in patients with chronic HCV infection than in healthy controls (42.6% vs. 4.7%; P<0.0001). Cryoglobulins were present in 68.4% of patients with HCV-related cirrhosis, which was nearly twice the figure in noncirrhotic HCV-infected patients and alcoholic cirrhotic patients. There were no differences in age, sex, aminotransferase levels or HCV genotype distribution in HCV-infected patients with or without cryoglobulinaemia. Only 13% of patients with chronic HCV infection and cryoglobulins showed symptoms of cryoglobulinaemia. There was a linear association between HCV RNA concentration in sera and in cryoprecipitates (P<0.0005). Patients with chronic HCV infection had a high prevalence of cryoglobulinaemia, especially in advanced forms of the disease, but clinical findings are few. There was no relationship with the genotype of HCV. The presence of HCV RNA in cryoprecipitates supported the hypothesis on the aetiological role of HCV in mixed cryoglobulinaemia.