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1.
目的 应用基因测序技术检测分析湖北地区慢性丙型肝炎患者的HCV基因型及其分布特点。方法 收集2013年2月~2015年7月武汉大学人民医院感染科447例HCV-RNA阳性的慢性丙型肝炎患者血浆标本,通过Sanger测序法测定HCV基因的NS5B区基因序列,然后与NCBI genebank数据库中HCV基因型数据进行比对,分析患者HCV基因型。结果 研究检测的慢性丙型肝炎患者血浆中,共检测出11种HCV基因型,分别是1a,1b,2a,3a,3b,6a,6b,1b/2a混合型,1b/2k混合型,6a/1b混合型和6d/6k混合型。其中,HCV1a型为7例(1.57%),1b型325例(72.71%),2a型67例(14.99%),3a型7例(1.57%),3b型20例(4.47%),6a型14例(3.13%),6b型2例(0.45%),1b/2a混合型2例(0.45%),1b/2k型1例(0.22%),6a/1b混合型1例(0.22%)和6d/6k混合型1例(0.22%)。结论 湖北地区慢性丙型肝炎患者的HCV-RNA基因型以1b型为主,其次为2a型,亦可见其他型别,提示湖北地区HCV流行的基因型呈多样性。  相似文献   

2.
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.  相似文献   

3.
徐辉  杨伟国  李永红 《国际检验医学杂志》2012,33(17):2081-2082,2084
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.  相似文献   

4.
目的探讨武汉市丙型肝炎病毒基因型分布特点。方法采用巢式PCR扩增c DNA、基因测序法检测武汉市区169份抗-HCV阳性和HCV RNA增高患者的HCV基因型,目标检测基因型为1a、1b、2a、2b、3a、3b、4、5a、5b、6a共10个型。结果武汉市区169例HCV感染者血清中,基因1型115例(68.04%)、基因2型32例(18.93%)、基因3型16例(9.47%)、基因6型6例(3.55%),各基因亚型例数和其所占比例依次为1 b型113例(66.86%)、2 a型31例(18.34%)、3b型12例(7.10%)、6a型6例(3.55%)、3a型4例(2.37%)、1a型2例(1.18%)、2b型1例(0.59%),未检出4型、5a、5b和混合基因型,169例患者HCV以基因1b型为主。结论武汉市区丙型肝炎感染HCV主要基因型为1b型,其次为2a、3b、6a,而3a、1a、2b比例极低。  相似文献   

5.
目的 应用基因测序技术检测分析武汉地区慢性丙型肝炎患者的HCV基因型及其分布特点.方法 收集2013年2月-2014年1月武汉大学人民医院感染科254例HCV-RNA阳性慢性丙型肝炎患者血浆标本,通过Sanger测序法测定靶片段的基因序列,然后与NCBI genebank数据库中HCV基因型数据进行比对,分析患者HCV基因型.结果 研究检测的慢性丙型肝炎患者血浆中,共检测出11种HCV基因型,分别是1a,1b,2a,3a,3b,6a,6b,1b/2a混合型,1b/2k混合型,6a/1b混合型和6d/6k混合型.其中,HCV1a型为2例(0.79%),1b型189例(74.41%),2a型35例(13.78%),3a型4例(1.57%),3b型12例(4.72%),6a型6例(2.36%),6b型1例(0.39%),1b/2a混合型2例(0.79%),1b/2k型1例(0.39%),6a/1b混合型型1例(0.39%)和6d/6k混合型1例(0.39%).结论 武汉地区HCV-RNA基因型以1b型为主,其次为2a型,亦可见其他型别,提示武汉地区HCV流行的基因型呈多样性.  相似文献   

6.
目的研究慢性丙型肝炎病毒感染者体内自身免疫抗体的动态变化,及其与HCV-RNA基因型的关系。方法 62例慢性(丙型病毒性肝炎患者HCV感染经RT-PCR证实。HCV基因型的检测采用Simmonds基因分型法。抗线粒体抗体(AMA)、抗肝肾微粒体抗体(LKM)、抗可溶性肝抗原(SLA)、抗去唾液酸糖蛋白受体抗体(ASG)和抗肝细胞膜抗原抗体(LMA)采用ELISA检测。结果 62例慢性丙型病毒性肝炎患者中,54例(87.75%)为Ⅰ型HCV感染,其中Ⅰa型4例(6.45%),Ⅰb型38例(61.27%),Ⅰc型12例(19.35%);4例(6.45%)为Ⅱa型;1例(1.63%)为混合型(Ⅰb型+Ⅱa型);2例(3.25%)为未分型。同时检测出Ⅰ型HCV感染的54例中有21例检出自身抗体阳性(38.9%),Ⅱa型HCV感染的4例中有1例检出自身抗体阳性(25%)。Ⅰb型和Ⅱa型混合感染的1例及未分型的2例均为阴性。62例HCV感染者中自身抗体阳性者共22例(35.48%),其中男7例(24.14%),女15例(44.45%),两者比较差异有统计学意义(P<0.05)。阳性患者平均年龄为(55.67±6.75)岁明显高于阴性患者(40.21±7.51)岁,差异有统计学意义(P<0.05)。结论在该地区的慢性丙型病毒性肝炎患者中,基因型分布以Ⅰb型为主。HCV感染者中普遍存在自身抗体,自身抗体的出现与年龄及性别有明显关系,但与HCV基因型无明显相关。基因Ⅰ型HCV感染者抗病毒治疗可诱发自身抗体水平的升高,但它是一过性的现象,在抗病毒治疗结束后部分自身抗体可降低甚至消失。  相似文献   

7.
目的 研究丙型肝炎患者HCV基因型的分布,探讨基因型在性别上的分布以及基因型与HCV RNA病毒载量的相关性.方法 收集2010年5-12月来自40家医院的206例丙型肝炎患者的血清标本,采用瑞士罗氏公司生产的定量PCR试剂(罗氏试剂)对进行HCV RNA检测,应用雅培公司生产的Abbott RealTime HCV GenotypeⅡ试剂(雅培试剂)对206例丙型肝炎患者的血清标本进行基因分型,分析基因型在性别上的分布以及HCV基因型与HCV RNA病毒载量的相关性.结果 206份HCV RNA阳性血清标本中HCV1型(未具体分1a和1b型)占3.4%(7/206)、1a型占1.0%(2/206)、1b型占59.7%(123/206)、2型占15.5%(32/206)、3型占13.1%(27/206)、6型占2.9%(6/206)、1/6混合型占2.4%(5/206)、2/4混合型占0.5%(1/206),未分型占1.5%(3/206).132例基因1型和65例非基因1型(2型、3型和6型)患者HCV基因型在性别上的分布差异无统计学意义(x2=0.000,P>0.05).188例患者不同基因型之间血清HCV RNA病毒载量差异有统计学意义(F=3.371,P<0.05).将197例HCV单基因型患者按地区分为东、南、西、北、中5组,基因型1型与非基因1型在地区分布上差异无统计学意义(x2=5.840,P>0.05).结论 丙型肝炎病毒感染以1b型为主,其次为基因2型.基因型在性别上的分布没有差异.基因1b型HCV RNA病毒载量高于基因3型,基因2型HCV RNA病毒载量高于基因3型,基因6型HCV RNA病毒载量高于基因3型.  相似文献   

8.
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.  相似文献   

9.
目的 人乳头瘤病毒(HPV)感染是宫颈上皮内瘤变及宫颈癌的主要致病原因.本文旨在探讨深圳市女性宫颈细胞中23种HPV感染的基因型分布情况.方法 从深圳市妇幼保健院体检中心的2 109例女性宫颈上皮细胞标本中提取23种HPV DNA,采用基因扩增及基因芯片技术对其宫颈细胞中的23种HPV基因型别进行检测,并对受检者的感染情况进行分析.结果 2 109例女性宫颈上皮细胞标本中检出HPV感染者446例,总的HPV感染率为21.15%(446/2 109),其中单一型别的阳性检出率为15.32%(323/2 109);单一型别的感染中HPV52型为47例,其阳性检出率为2.23%(47/2 109),其次是HPV43和58型均为38例,其阳性检出率均为1.80%(38/2 109),是单一型别感染中最主要的型别.混合型HPV感染123例,其阳性检出率为5.83%(123/2 109);其中HPV16+43、HPV16+58型各5例,均占混合型感染的4.07%(5/123),其次是HPV43+52、HPV43+56、HPV43+66型的二重感染各3例,均占混合型感染的2.44%(3/123),是混合型感染的主要型别.结论 HPV43、HPV52、HPV58单一型别及HPV16+43和HPV16+58型是感染深圳市女性宫颈细胞的主要基因型别,基因芯片检测技术可应用于宫颈细胞标本,一次可检测23种HPV基因型别,特异性强,敏感性高,对中国女性宫颈HPV感染基因型的分子流行病学的调查研究具有重要的意义.  相似文献   

10.
目的:了解芯片技术对南昌地区HIV-HCV合并感染者的HCV基因型分布。方法2008年10月至2013年11月期间,在南昌地区就诊的285例HIV感染患者进行HCV筛查,并对HCV阳性患者的基因型进行芯片技术检测,将其检测结果与测序结果进行比对;同时对HIV-HCV合并感染者进行流行病学调查。结果芯片技术可检测出约900拷贝/ml的丙型肝炎病毒的基因型,HIV感染者中,HCV阳性比率为19.3%,HIV-HCV合并感染者的HCV基因型分布情况为1b型41例,2a型9例;3b型1例,6型1例,还有1 b+2a型3例;芯片检测结果与基因测序结果完全一致。结论芯片技术灵敏度高,准确性好,HCV基因型分布以1b基因型为主,其次为2a型;HIV感染者的感染传播途径有了新变化,性传播感染HIV比例在增加,但性传播合并感染的比例较低。  相似文献   

11.
Multiple infection by different hepatitis C virus (HCV) genotypes may be of great clinico-pathologic interest. In this study we determined the effective prevalence of coinfections by two or more HCV genotypes in 213 subjects with HCV-positive chronic hepatitis by using genotype-specific polymerase chain reaction (PCR), genotype-specific probe hybridization, and direct sequencing. The most prevalent genotype was HCV-1b (54%). HCV-2 (a/c) was also prevalent (27%), and types 1a and 3a were found in 5% and 3% of patients, respectively. A mixed infection was detected in 23 patients (10.8%): 4 out of 23 were coinfected by types 1a + 1b, while the remaining 19 patients had a b + 2 (a/c) mixed infection. Further analysis based on restriction fragment length polymorphism (RFLP) on type-specific PCR products was used to verify genotyping results. Only four coinfections (1a + 1b in 2 patients and 1b + 2 (a/c) in the remaining 2 patients, respectively) were confirmed by enzyme cleavage. All patients with true coinfection had long-lasting infection and liver cirrhosis. Both true and false mixed infections resulting from RFLP analysis were confirmed by direct sequencing of type-specific amplification products. We also determined a recurrent C/T transversion at position 618 in all sequenced samples. In 4 cases another point mutation (G/A at position 626) was found, reducing the number of mismatches between HCV-2 and HCV-1b from 4 to 3 (or 2). Interestingly, all HCV-2 isolates sequenced showed the highest degree of nucleotide homology with HCV-2 subtype c, confirming the relatively high prevalence of this subtype in Italy. In conclusion, we showed the possibility of multiple infection by different HCV types in the general population of chronically infected patients without particular risk factors, even if in a low percentage of cases. Further studies are needed to assess the clinical relevance of chronic HCV infection with multiple genotypes.  相似文献   

12.
There is no comprehensive study available on the natural hepatitis C virus (HCV) polymorphism in sites associated with resistance including all viral genotypes which may present variable susceptibilities to particular direct-acting antivirals (DAAs). This study aimed to analyze the frequencies, genetic barriers, and evolutionary histories of naturally occurring resistance-associated variants (RAVs) in the six main HCV genotypes. A comprehensive analysis of up to 103 RAVs was performed in 2,901, 2,216, and 1,344 HCV isolates for the NS3, NS5A, and NS5B genes, respectively. We report significant intergenotypic differences in the frequencies of natural RAVs for these three HCV genes. In addition, we found a low genetic barrier for the generation of new RAVs, irrespective of the viral genotype. Furthermore, in 1,126 HCV genomes, including sequences spanning the three genes, haplotype analysis revealed a remarkably high frequency of viruses carrying more than one natural RAV to DAAs (53% of HCV-1a, 28.5% of HCV-1b, 67.1% of HCV-6, and 100% of genotype 2, 3, 4, and 5 haplotypes). With the exception of HCV-1a, the most prevalent haplotypes showed RAVs in at least two different viral genes. Finally, evolutionary analyses revealed that, while most natural RAVs appeared recently, others have been efficiently transmitted over time and cluster in well-supported clades. In summary, and despite the observed high efficacy of DAA-based regimens, we show that naturally occurring RAVs are common in all HCV genotypes and that there is an overall low genetic barrier for the selection of resistance mutations. There is a need for natural DAA resistance profiling specific for each HCV genotype.  相似文献   

13.
Etiologic variations of hepatocellular carcinoma (HCC) exist in different geographic areas of the world. Hepatitis B virus infection is associated with HCC. However, hepatitis C virus (HCV) infection plays an increasingly more important role in the development of HCC and is associated with more than 30% of HCC in Taiwan. The prevalence of HCV infection and HCV genotypes vary in different geographic areas. The prevalence of HCV genotype 1b (HCV-1b) was around 50-70% in Taiwan and even varied in different townships. In addition to host factors, HCV genotypes may be associated with the development of HCC. In our study, the prevalence of HCV-1b in patients with HCC was significantly higher than in those with liver cirrhosis and chronic hepatitis; multivariate analysis revealed that the disease severity was significantly correlated with age and HCV-1b. Furthermore, HCV-1b was associated with a lower response rate to interferon (IFN) therapy than HCV-2. Our study has demonstrated that mutations in the IFN sensitivity-determining region, spanning nucleotides 2,209-2,248 in the NS5A region, correlate with the sustained virological response to combination therapy with IFN and ribavirin in patients with chronic HCV-1b infection in Taiwan. A third-generation enzyme immunoassay for antibody to HCV can be used to predict viremia and monitor the virological response.  相似文献   

14.
GSK2485852 (referred to here as GSK5852) is a hepatitis C virus (HCV) NS5B polymerase inhibitor with 50% effective concentrations (EC50s) in the low nanomolar range in the genotype 1 and 2 subgenomic replicon system as well as the infectious HCV cell culture system. We have characterized the antiviral activity of GSK5852 using chimeric replicon systems with NS5B genes from additional genotypes as well as NS5B sequences from clinical isolates of patients infected with HCV of genotypes 1a and 1b. The inhibitory activity of GSK5852 remained unchanged in these intergenotypic and intragenotypic replicon systems. GSK5852 furthermore displays an excellent resistance profile and shows a <5-fold potency loss across the clinically important NS5B resistance mutations P495L, M423T, C316Y, and Y448H. Testing of a diverse mutant panel also revealed a lack of cross-resistance against known resistance mutations in other viral proteins. Data from both the newer 454 sequencing method and traditional population sequencing showed a pattern of mutations arising in the NS5B RNA-dependent RNA polymerase in replicon cells exposed to GSK5852. GSK5852 was more potent than HCV-796, an earlier inhibitor in this class, and showed greater reductions in HCV RNA during long-term treatment of replicons. GSK5852 is similar to HCV-796 in its activity against multiple genotypes, but its superior resistance profile suggests that it could be an attractive component of an all-oral regimen for treating HCV.  相似文献   

15.
BACKGROUND: Formation of transmembrane ion channels by hepatitis C virus (HCV) p7 and abrogation of channel function by amantadine was demonstrated in vitro. The relevance of HCV p7 amino acid (aa) variations for response to antiviral therapy with amantadine is unknown. METHODS: HCV p7 was sequenced in 86 individuals who were infected with HCV genotype 1. Thirty-six of 86 patients received amantadine within an interferon-alpha (IFN-alpha)-based antiviral therapy. Helical wheel modelling for HCV p7 was performed. RESULTS: No significant correlation of overall aa variations within HCV p7 was observed with response to IFN-alpha-based therapy with amantadine in HCV genotype 1alpha/b infected patients. When analysis was restricted to non-conservative aa variations, a higher number of aa substitutions within complete HCV p7 and transmembrane helix 2 was associated with non-response in HCV-1b-infected patients receiving therapy with amantadine (P=0.015 and P=0.037, respectively), without amantadine (P=0.106 and P=0.118, respectively), and in the total cohort of HCV-1b-infected patients (P=0.00007 and P=0.011, respectively). Furthermore, substitution L20F was observed more often in non-responders than responders with HCV-1b infection and therapy with amantadine (P=0.099). By in silico modelling, aa 20 was located toward the p7 channel lumen. Substitution L20F may impair amantadine action by altering the shape of the ion channel pore. CONCLUSION: Substitution L20F within HCV p7 may be associated with non-response to combination therapy specifically with amantadine in HCV-1b-infected patients. Non-responders with HCV-1b infection showed higher numbers of non-conservative aa variations within HCV p7 than responders, irrespective of the application of amantadine.  相似文献   

16.
BACKGROUND: The TT virus (TTV) is a newly identified blood-borne virus. Its association with disease is still unknown, and screening of blood donors has not been implemented. Several genotypes of the TTV have been identified. STUDY DESIGN AND METHODS: Three hundred seventy healthy blood donors were randomly selected and tested for TTV by the PCR method. Sequencing of a part of the genome was performed to identify various genotypes of the virus. ALT levels were determined in both infected and uninfected individuals. RESULTS: The TT virus (TTV), was detected in the sera of 23 (6.2%) of 370 healthy Icelandic blood donors; this prevalence is lower than that reported in Japan but higher than that in Scotland. The virus was found in all groups over the age of 19. Sequencing and phylogenetic analysis of 202 bp from open reading frame 1 demonstrated genotypes 1b and 2b 2c and genotype 4 isolates, with the latter bearing 89-percent nucleotide homology with other genotype 4 sequences deposited at GenBank. One sample showed a mixed genotype 1b/2c infection. Serum ALT levels were within normal limits in all infected individuals. CONCLUSION: The TTV carrier state does not cause significant liver injury.  相似文献   

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