肝癌患者血清中乙型肝炎病毒基因分型及临床意义

目的:探讨血清中乙型肝炎病毒(HBV)基因型及HBV DNA水平与肝细胞癌的关系。方法:应用巢式聚合酶链反应扩增乙型肝炎病毒与基因,用末端标记方法对PCR产物标记并直接测序,测序结果和GenBank中登录的标准基因型序列相比较,应用荧光定量PCR法检测HBV DNA水平。对61例肝癌、65例慢性乙型肝炎、10例乙型肝炎病毒携带者进行了检测。结果:136例中B基因型59例(43.4%)、C基因型77例(56.6%),随着病情加重,C基因型比例逐渐增高;不同基因型HBV感染的肝癌患者间HBV DNA水平差异有显著意义,P<0.05;在慢性乙型肝炎患者中,HBV DNA水平差异无显著性意义。结论:本地区乙型肝炎病毒以B、C基因型为主,乙型肝炎病毒C基因型及高水平的HBV DNA感染与肝癌的发生相关。     

湖北地区乙型肝炎病毒基因型分布与临床的相关性

目的 了解湖北地区乙型肝炎病毒(HBV)基因型的分布及其与临床的相关性。 方法 选择湖北地区HBV DNA阳性的慢性HBV感染者190例,其中乙型肝炎表面抗原(HBsAg)携带者52例、慢性乙型肝炎56例、重型肝炎32例、肝硬化22例、原发性肝癌28例,应用多对型特异性引物-聚合酶链反应检测HBV的基因型。 结果 多对HBV型特异性引物法可快速准确鉴定HBV的基因型。190份HBV DNA阳性血清标本中,B基因型140例(73.7％),C基因型42例(22.1％),BC混合型8例(4.2％),未发现A、D和E基因型;B基因型在重型肝炎和肝癌患者中占绝对优势,分别为87.5％和89.3％,显著高于HBsAg携带者的67.3％;B基因型患者血清丙氨酸氨基转移酶水平(253.1±306.7)U／L高于C基因型患者的(154.1±192.9)U／L,差异有统计学意义(P<0.05);除HBsAg携带者外的慢性HBV感染者中,B基因型患者血清抗-HBe阳性率50.5％(53／105)显著高于C基因型的18.5％(5／27),P<0.01。 结论 多对型特异性引物同时进行聚合酶链反应的基因分型方法可用于HBV基因型的流行病学调查;湖北地区存在HBV的B、C和BC混合基因型,B型为本地区的优势基因型并在严重肝病和肝癌中的比例较高,基因型的分布可能有较大的地区差异。     

慢性乙型肝炎拉米夫定耐药患者HBV基因型及ALT变化的观察

观察慢性乙型肝炎患者用拉米夫定治疗后HBVP基因变异与不同HBV基因型感染及HBV DNA复升水平和转氨酶变化.收集51例慢性乙型肝炎患者用拉米夫定治疗52-78周后发生YMDD变异的血清标本,对照组128例未用拉米夫定治疗的慢性乙型肝炎患者血清标本,应用聚合酶链反应方法,测定HBV DNA基因型;用限制性片段长度多态性分析方法(PCR RELP)测定HBV DNA YMDD变异;同时进行HBV DNA定量分析.结果显示51例拉米夫定治疗后HBV DNA基因变异患者以B型和C型为主,分别为10例(19.6%)和39例(76.47%),B C混和型2例(3.92%),未见其它基因型.拉米夫定治疗引起HBVDNAYMDD变异可以发生在不同HBV基因型感染的慢性乙型肝炎患者中,与对照组比较二者没有显著性差异.     

厦门市乙型肝炎患者病毒基因型的分析

目的:采用多对型特异性引物,通过巢式PCR法检测厦门市乙型肝炎患者血清中乙型肝炎病毒(HBV)基因型的分布情况.方法:收集250例HBV感染患者血清,提取血清中HBV DNA作为模板,设计HBV前S1基因和S基因中区域内设计出10条内外引物,并将其中8条型特异性内引物分成A,B两组,分别扩增A,B,C和D,E,F型HBV,然后将第2轮PCR产物以用30g/L琼脂糖进行电泳,根据PCR产物电泳显示的产物长度判定HBV基因型,以了解厦门HBV基因型分布情况.结果:共120例确定了HBV基因型.患者群中慢性乙型肝炎90例,占75.0%,急性乙型肝炎、肝炎肝硬化、原发性肝癌分别占5.8%(7/120)、6.7%(8/120)和12.5%(15/120).分型结果:B型58例(48.3%)、C型30例(25.0%),B/C混合型32例(26.7%).HBeAg阳性患者中B基因型占63.8%,B/C型混合感染21.9%;抗-HBe阳性患者中以B/C型混合感染68.8%,B型25.9%,HBeAg阳性组与抗-HBe组之间比较发现B型和B/C混合型之间(P<0.05).结论:厦门乙型肝炎患者HBV基因型以B型为主,B/C混合感染是一个值得重视的问题.     

慢性乙型肝炎患者HBV基因型和亚型分布调查

目的研究慢性乙型肝炎患者HBV基因型和亚型流行情况。方法应用HBV基因型和亚型特异性引物PCR法对北京、长春、大连、西安、石家庄、郑州和合肥7个城市660份HBVDNA阳性慢性乙型肝炎患者血清进行基因型和亚型分析。结果在660份HBVDNA阳性血清中,B基因型、C基因型和B/C混合感染分别为16.67%(110/660)、74.54%(492/660)和8.79%(58/660);在C基因型中,C1亚型6例(1.22%)、C2亚型473例(96.14%)、C1/C2混合基因亚型13例(2.64%);B基因型均为Ba亚型,B基因型和C基因型混合感染者均为Ba与C2亚型混合感染,未发现其他基因型和基因亚型;不同基因型感染患者HBeAg阳性率差异无统计学意义(P=0.153);B基因型和C基因型患者之间血清HBVDNA水平差异无统计学意义(6.37±1.62lgcopies/ml对6.29±1.76lgcopies/ml),但均高于B和C基因型混合感染患者(5.25±1.65lgcopies/ml)。结论这7个城市慢性乙型肝炎患者以B基因型和C基因型感染为主,有部分B/C基因型混合感染。HBV亚型以Ba和C2亚型占优势。     

替诺福韦治疗不同HBV基因型慢性乙型肝炎患者疗效研究*

目的 探讨替诺福韦治疗不同HBV基因型感染的慢性乙型肝炎(CHB)患者的疗效和血清金属硫蛋白(MT)、白细胞介素29(IL-29)和外周血淋巴细胞程序性死亡受体-1(PD-1)表达的变化。方法 2017年1月～2020年11月我院诊治的CHB患者119例,均接受替诺福韦治疗。采用ELISA法检测血清MT和IL-29水平,使用流式细胞仪检测外周血T 淋巴细胞PD-1表达水平。结果 在本组119例CHB患者中,检出HBV B基因型31例(26.1%),C基因型77例(64.7%),B/C 混合基因型11例(9.2%);基线资料比较,B基因型CHB患者血清ALT和AST水平分别为(154.1±46.7)U/L和(83.3±26.8)U/L,显著高于C基因型患者【分别为(135.8±40.3)U/L和(68.5±20.6)U/L,P＜0.05】或B/C混合基因型患者【分别为(138.9±50.2)U/L和(71.6±23.9)U/L,P＜0.05】,而血清HBV DNA水平为(6.7±1.1)lg copies/ml,显著低于C基因型感染患者【(7.8±1.4)lg copies/ml,P＜0.05】或B/C混合型感染者【(7.4±1.0)lg copies/ml,P＜0.05】;在治疗24 w和48 w末,三组血清ALT复常率无显著性差异(P>0.05),而B基因型和C基因型感染患者血清HBV DNA阴转率分别为96.8%和96.8%,和88.3%和93.5%,均显著高于B/C混合型感染患者(分别为63.6%和81.8%,P<0.05);在治疗48 w,B基因型CHB患者血清MT水平显著高于C基因型或B/C基因型(P＜0.05),血清IL-29水平显著高于C基因型(P＜0.05),而外周血CD4⁺和CD8⁺T淋巴细胞表面PD-1表达水平显著低于C基因型或B/C混合基因型患者(P＜0.05)。结论 不同HBV基因型感染的CHB患者可能对替诺福韦治疗的疗效存在差异,深入了解这些差异可能对研究不同基因型病毒感染患者临床转归有帮助。     

乙型肝炎病毒基因分型及其临床意义

目的了解慢性乙型肝炎病毒(HBV)感染者HBV基因分型及其对慢性肝病的影响,为制定针对不同HBV基因型抗病毒的个体化方案提供分子病毒学依据。方法临床确诊的慢性乙型肝炎、乙型肝炎肝硬化及肝癌患者314例,采用RDB法对HBV进行基因分型检测。结果山西地区的200例慢性乙型肝炎患者所感染的HBV均为B和C基因型,分别占56%、26%,并存在混合感染(17%);C与B基因型患者相比,血清病毒载量高、肝脏损伤严重;混合感染的患者与单一基因型感染者相比病毒载量更高、肝损伤更严重;肝硬化患者感染的HBV主要为C基因型及B、C混合感染,且肝损害严重、病毒复制率高;肝癌患者中C基因型感染占42.19%,B、C混合感染占37.5%,B基因型感染可能与年轻患者肝癌的发生有关。结论B基因型HBV感染与C基因型及混合感染相比,病毒载量低、肝损害轻,但年轻患者应监测肝癌的发生;C基因型及混合感染的患者预后较差,肝硬化、肝癌发生率高,应进行积极有效的治疗,防止严重肝病发生。     

安康地区乙型肝炎病毒基因型分布及临床相关性研究

目的:探讨安康地区HBV基因型的分布状况及其与临床的相关性。方法:应用型特异性引物法检测陕西省安康地区1275例HBV感染者的HBV基因型,并分析了287例住院患者的基因型与临床的相关性。结果:1275例HBV感染者中B基因型902例(70.75%),C基因型370例(29.02%),B/C混合型3例(0.23%),未检测到其他基因型。287例住院患者基因型分布在性别上无明显差异(P=0.32),B型、C型患者在携带者、慢性肝炎、重型肝炎、肝硬化和肝癌中的分布无明显差异(P=0.45),在ALT水平及HBV DNA载量上无明显差异(分别为P=0.75,P=0.63);C基因型患者的HBeAg阳性率(70.37%)明显高于B基因型患者(44.17%),P0.001。结论:安康地区HBV感染者基因型以B型为主,C型次之,C基因型患者的HBeAg阳性率明显高于B基因型患者,B、C基因型在性别、疾病状态、肝脏炎症活动度及ALT水平上无显著性差异。     

浙南地区HBV基因型分布特征及其与慢性乙型肝炎患者血清HBV DNA水平和HBeAg表达的关系

目的分析浙南地区慢性乙型肝炎患者HBV基因型分布特征,探讨不同基因型与患者血清中HBV DNA水平及HBeAg表达的关系。方法收集2011年6月-2012年3月161例就诊的HBV DNA阳性患者血清标本,采用型特异性引物PCR方法对HBV进行基因分型并用实时荧光定量PCR方法测量HBV DNA含量(拷贝/ml)和ELISA方法检测HBeAg;对HBV DNA含量进行对数转换使其符合正态分布后,运用χ2检验分析基因型与HBeAg阳性率的关系,t检验分析B、C基因型患者HBV DNA水平的差异。结果 161例患者中,B型41例,占25.5%;C型118例,占73.3%;BC混合型2例,占1.2%。C型患者HBV DNA水平和HBeAg阳性率分别为(5.84±1.40)log10拷贝/ml和64.4%,高于B型的(5.49±1.33)log10拷贝/ml和53.7%,但差异均无统计学意义(P0.05)。结论浙南地区HBV基因型以C型为主,B型次之。C基因型HBV DNA水平与HBeAg阳性率均高于B基因型,肝硬化的发生与C基因型HBV密切相关。     

365例乙型肝炎基因型分布与临床相关性研究

目的:探讨湖北地区乙型肝炎病毒(HBV)基因型分布情况以及其与临床的相关性.方法:选取湖北地区HBV DNA阳性的慢性乙型肝炎患者365名,应用多对型特异性引物聚合酶链反应检测HBV的基因型.结果:在365份HBV DNA阳性血清中,除3份标本不能分型外,B型占78.45%,C型和BC混合型占17.68%和3.87%,未发现其他HBV基因型.B型患者血清ALT水平显著高于C型和BC混合型患者(P=0.01,P＜0.001),血清TBil、HBeAg阳性率在各基因型患者间差异无显著性意义.慢性肝炎患者中B基因型显著多于C基因型(P=0.012),肝硬化及肝癌患者中,B、C基因型分布的差异有显著性意义(P=0.007,P=0.001),重症肝炎患者的基因型分布无显著性意义.结论:湖北地区存在B、C、BC混合3种HBV基因型,且以B型为优势基因型.在慢性肝炎患者中B型比例较高,肝硬化及肝细胞癌患者中C型比例较高.     

慢性乙型肝炎和肝硬化患者血清HBV基因型分析

目的 分析慢性乙型肝炎和肝硬化患者血清乙型肝炎病毒(HBV)分型分布情况。方法 2015年6月～2018年5月南京中医药大学附属南京市第二医院就诊的慢性乙型肝炎患者261例,乙型肝炎肝硬化患者30例,肝细胞癌4例,采用测序法检测血清HBV基因型。结果 在295例HBV感染者中,有132例(44.7%)为B型感染,161例(54.6%)为C型感染,2例(0.7%)为D型感染;慢性乙型肝炎患者与肝硬化患者血清TBIL、ALT和AST水平比较差异均无统计学意义(P>0.05);肝硬化患者血清肝纤维化指标(P<0.05)、血清HBV DNA载量(P<0.05)和血清HBeAg阳性率(x²=5.798,P<0.05)均显著高于慢性乙型肝炎患者;乙型肝炎肝硬化患者和肝细胞癌患者C型感染比例均显著高于慢性乙型肝炎患者,差异具有统计学意义(P<0.05)结论 慢性乙型肝炎和肝硬化患者HBV感染以B基因型和C基因型为主,而肝硬化患者以C型感染居多,提示C型感染患者可能比B型患者更容易出现严重的肝损伤,并产生严重的临床结局。     

Hepatitis B virus genotypes and clinical manifestation among hepatitis B carriers in Thailand

BACKGROUND: Hepatitis B virus (HBV) genotypes have distinct geographic distributions. The aim of the present study was to evaluate the distribution of HBV genotypes and their clinical relevance in Thailand. METHODS: Hepatitis B virus genotypes among 107 hepatitis B carriers residing in Thailand were evaluated using serologic and genetic methods. They were clinically classified into asymptomatic carriers with normal serum alanine transaminase (ALT) levels and patients with chronic liver disease, such as those with chronic hepatitis (CH), liver cirrhosis (LC) and hepatocellular carcinoma (HCC). RESULTS: Hepatitis B virus genotype distribution among the 107 patients was 25.2% for genotype B, 72.0% for genotype C and 2.8% for genotype D. The serum ALT levels, HBV-DNA and hepatitis B e antigen positivity were significantly higher in carriers infected with genotype C HBV than in those infected with genotype B (P < 0.05). The proportion of genotype B HBV was higher in asymptomatic carriers than in patients with CH and those who developed liver disease, such as LC and HCC (45.5, 16.9 and 25.0%, respectively; P < 0.05). In contrast, the proportion of genotype C HBV was higher in patients who developed liver disease and CH than in asymptomatic carriers (68.7, 83.0 and 50.0%, respectively; P < 0.05). Phylogenetic analysis based on entire genome sequences revealed three HBV isolates, which were classified into a subgroup of genotype C in isolates from South-East Asian countries. CONCLUSIONS: Genotypes B and C are the predominant types among hepatitis B carriers residing in Thailand and those genotypes influence the clinical manifestation in carriers with chronic hepatitis B infection.     

Hepatitis B virus genotypes and hepatocellular carcinoma in Thailand

AIM: The role of hepatitis B virus (HBV) genotypes on the clinical features and prognosis of patients with hepatocellular carcinoma (HCC) is currently unknown. The aim of the present study was to evaluate the distribution of HBV genotypes and their clinical relevance in Thai patients. METHODS: HBV genotypes were determined by PCR-RFLP in stored sera of 93 asymptomatic carriers, 103 patients with chronic hepatitis, 60 patients with cirrhosis and 76 patients with HCC. The clinical data were analyzed in relation to the HBV genotype. RESULTS: HBV genotypes C and B were predominant in Thailand, accounting for 73% and 21%, respectively. The distributions of genotypes B and C were similar in HCC patients compared to the other groups. Genotype C was significantly more common in HCC patients who were under 40 years old than genotype B (18% vs 0%, P= 0.03), but was significantly less common in patients older than 60 years (26% vs 56.5%, P= 0.01). The positive rate of hepatitis B e antigen (HBeAg) in patients with genotype C was significantly higher than that in patients with genotype B (71.6% vs 44.4%, P= 0.03 in chronic hepatitis; 56.8% vs 11.1%, P= 0.01 in cirrhosis). There were no differences between HCC patients with genotypes B and C regarding tumor staging by CLIP criteria and the overall median survival. Multivariate analyses showed that HBV genotype was not an independent prognostic factor of survival in HCC patients. CONCLUSION: Patients with genotype C had a higher positive rate of HBeAg and exhibited earlier progression of cirrhosis and HCC than those with genotype B. However, there were no differences in the risk of developing HCC and its prognosis between patients with these genotypes.     

Hepatitis B viral genotype in Taiwanese patients with acute hepatitis B

BACKGROUND/AIMS: The correlation of HBV genotype with clinical outcome has been recognized in chronic hepatitis B patients. However, there are few reports on the distribution and clinical significance of HBV genotypes in acute hepatitis B patients. METHODOLOGY: Nineteen acute hepatitis B patients were identified and their HBV genotypes were determined. The serological and clinical data were thus compared between patients with different HBV genotypes. RESULTS: Two HBV genotypes (B and C) were found in the patients. Genotype B was more predominant than genotype C (12 vs. 7). The age, serum alanine aminotransferase level, serum alpha-fetoprotein level, and serum HBV DNA level were not significantly different between patients infected with genotype B or C. None of them had persistent HBsAg for longer than 6 months. CONCLUSIONS: Genotype B predominates in acute hepatitis B patients in Taiwan; however, the clinical features between genotype B and genotype C patients are comparable.     

Hepatitis B genotypes and the response to interferon therapy

BACKGROUND/AIMS: Possible pathogenic differences among hepatitis B virus (HBV) genotypes have been observed; however, the response to interferon therapy among HBV genotypes remains unknown. We therefore analyzed the efficacy of interferon alfa in the treatment of chronic hepatitis B patients with different HBV genotypes. METHODS: Fifty-eight genotype B or C infected chronic hepatitis B patients who had been treated with interferon alfa-2b were retrospectively studied. The response to interferon was defined as normalization of serum aminotransferase level, loss of hepatitis B e antigen and HBV DNA 48 weeks post-treatment. RESULTS: Baseline data of both groups of patients were comparable; however, genotype C patients had a higher serum aminotransferase level and a higher frequency of core promoter mutation. The response rate was 41% and 15% in genotype B and C patients, respectively (p=0.045). In those with higher serum aminotransferase levels, the response rate was 50% and 17%, respectively (p=0.025). Additionally, younger age and genotype B infection may predict a better response to interferon alfa. CONCLUSIONS: HBV genotype C, compared to genotype B, is associated with a higher frequency of core promoter mutation, and a lower response rate to interferon alfa therapy.     

HBV genotypes in India: do they influence disease severity?

Aims:  Association of HBV genotypes (especially A and D) with severity of liver disease is controversial. We studied the influence of HBV genotypes on liver disease severity among Indian patients.
Methods:  We selected 247 HBV infected patients (42 acute hepatitis, 87 carriers, 44 chronic hepatitis B [CHB], 35 liver cirrhosis [LC] and 40 hepatocellular carcinoma [HCC]). Genotyping of stored sera was performed using genotype-specific enzyme-linked immunosorbent assay (ELISA) and restriction fragment length polymorphism (RFLP). The distribution of genotypes in disease states of differing clinical, histological and biochemical severity were compared.
Results:  The most common genotype was D (162/237, 68.3%), followed by A (61, 25.7%) and C (14, 5.9%). The distribution of HBV genotypes between patients with acute hepatitis and CHB (carriers + CHB + LC + HCC), or between carriers and disease states (CHB + LC + HCC), or between mild chronic infection (carriers + CHB) and complications of chronic HBV infection (LC + HCC) was similar. Eighty-seven patients had liver biopsy; the median histological activity index (HAI) and fibrosis stage at baseline were similar between genotype groups (four [1–9] genotype A [ n  = 28]), three (2–4) genotype C ( n  = 4) and four (1–10) genotype D ( n  = 55); P  = 0.33 for HAI score; (0.5 [0–6] genotype A, 0.5 [0–4] genotype C and 1 [0–6] genotype D; P  = 0.92 for fibrosis stage). The response to therapy was similar between the genotypes.
Conclusion:  Clinical, histological severity and therapeutic responses are similar among patients with HBV genotypes A and D.     

拉米夫定治疗无良好应答患者乙型肝炎病毒P区突变与基因型的关系

目的 观察拉米夫定治疗后无良好应答的慢性乙型肝炎患者HBV P区变异情况与基因型的关系.方法 对631例拉米夫定治疗后无良好应答的慢性乙型肝炎患者进行研究.通过荧光定量PCR或核酸测序确定HBV基因型,直接测序观察P区突变,实时荧光定量PCR方法检测患者病毒载量,比较不同基因型患者的HBV DNA水平及HBV P区变异情况.计量资料采用成组设计资料t检验,计数资料采用x~2检验或Fisher精确检验.结果 631例慢性乙型肝炎患者中,B基因型HBV感染者272例,C基因型感染者359例,C基因型感染者患者年龄为(39.1±11.4)岁,明显大于B基因型感染患者的(33.7±9.7)岁(t=-6.55,P<0.01).C基因患者病毒载量为(5.96±1.22)log_(10)拷贝/ml,高于B基因型患者的(5.58±1.21)log_(10)拷贝/ml,t=-2.01,P<0.05.A181V/T变异在C基因型的发生率高于B基因型(0.4%比5.3%,χ~2=12.23,P<0.01),M204I/V,L180M、T184A/G/I/S、S202G/I和V173L变异发生率在B、C基因型之间差异无统计学意义(P值均>0.05).M204I在B基因型的发生率为20.6%,高于C基因型的13.9%(χ~2=4.91,P<0.05);M204V和M201Ⅳ变异在B、C基因型中的发生率差异无统计学意义(χ~2值分别为1.70和2.21,P值均>0.05).拉米夫定耐药发生率在B、C基因型间差异无统计学意义(χ~2=0.00,P>0.05).结论 拉米夫定常见耐药位点在B、C基因型之间无明显差异,但是C基因HBV感染患者病毒载量高于B基因型HBV感染患者;M204I变异在B基因型中出现频率高于C基因型,拉米夫定加用或改用阿德福韦酯后可能会使A181V/T变异在C基因型出现的概率高于B基因型;年龄、免疫因素和非常见位点的变异或许是影响拉米夫定疗效的重要因素.