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目的建立重型乙型肝炎患者血清乙型肝炎病毒(HBV)DNA克隆并测序,从全基因水平分析HBV基因变异与重型乙型肝炎发病的关系。方法10例重型乙型肝炎患者血清提取HBV DNA,聚合酶链反应(PCR)扩增HBV全基因。PCR产物构建到PUCm-T载体上,转化至大肠杆菌感受态DH-5α细胞,经酶切鉴定,获得含3.2Kb HBVDNA的重组克隆菌,全基因测序,分析各读码框核苷酸和氨基酸变化。结果4例成功构建HBV DNA克隆,并完成全基因测序。其中3例在前C区发生G1896A变异,产生一个终止密码子,导致HBeAg缺失;1例在C启动子区1762、1764双位点出现突变;有多处点突变及缺失变异分布于PreS2区及C区已知细胞毒T淋巴细胞、B淋巴细胞和T淋巴细胞的细胞表位。结论该法可用于临床研究HBV病毒基因结构与重型乙型肝炎发病的关系,并为进一步研究其HBV基因功能奠定基础。 相似文献
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慢性乙型肝炎患者血清及肝组织中乙型肝炎病毒核心基因的研究 总被引:1,自引:0,他引:1
36例肝穿刺诊断为慢性乙型肝炎患者的肝组织及血清,用HBV全基因核酸杂交法及聚合酶链反应(PCR)法扩增了病毒的PreC/C区基因片段。发现PCR法扩增后再经Southern吸印转移及核酸杂交(PCR-SBH)可显著提高检出HBV基因的敏感性。对5例无任何血清HBV标记者证实在其肝内存在HBV基因。用AvaⅡ,sau3AI,XmnI,BstNI及TaqI酶切图谱分析上述病例的c基因,与pADR-1C基因比较,有4例(5份标本)有Sau3AI的异常酶切位点,提示HBV不仅PreC区存在变异株,C基因中亦可有变异。 相似文献
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乙型肝炎病毒基因型S基因PCR-RFLP分型方法的建立 总被引:57,自引:1,他引:56
目的 建立乙型肝炎病毒(HBV)S基因聚合酶链反应-限制性片段长度多态性(PCR-RFLP)的基因型分型方法并验证其准确性。方法 比较GenBank中HBV223株各基因型全序列的S基因氨基酸及核苷酸序列,设计限制性内切酶BsrⅠ、StyⅠ、DpnⅠ和HpaⅡ鉴别HBVA-F基因型的方法,选择经前SPCR-PFLP的方法已鉴定出的HBVB、C和D基因型标本各30例,用此方法鉴定验证,并随机对HBVB、C和D基因型各3株标本进行S基因测序证实。结果 两种PCR-RFLP及S基因测序鉴定HBV基因型结果一致。结论 S基因PCR-RFLP的方法能够准确鉴定HBV基因型,此方法灵敏性高,特异性强,且酶切图谱简明单一,易于识别,适宜于HBV基因型的流行病学调查。 相似文献
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乙型肝炎病毒逆向点杂交基因分型法建立及应用 总被引:4,自引:0,他引:4
目的 利用逆向点杂交技术建立乙型肝炎病毒(HBV)基因分型新方法,通过对HBV DNA阳性血清抽样标本进行基因分型,了解佛山地区HBV基因型分布状态。方法 以HBV基因组的X区序列为主设计分型引物与探针,将活性氨基标记的探针依次固定在尼龙膜上,制成检测膜条。用标记生物素的引物进行HBV DNA扩增,将扩增产物与检测膜条杂交,以POD与TMB显色,判断基因分型结果,通过与基因测序结果比较确定新方法的有效性。从佛山地区HBVDNA阳性患者血清中随机抽取300份,用新建方法进行HBV基因分型检测。结果 新建HBV逆向点杂交基因分型方法可对拷贝数在10^3~10^9/ml之间的300份HBV DNA阳性抽检血清进行基因分型,发现B型147例,占49.0%;C型136例,占45.3%;D型1例,占0.3%;B、C混合型12例,占4.0%;C、D混合型4例,占1.3%;未发现A、E和F型。新方法基因分型结果与测序结果一致。结论 利用逆向点杂交技术可以准确有效和简便经济的进行HBV基因分型,适用于临床检测与流行病学研究;在佛山地区,人群中感染的HBV以B、C型为主。 相似文献
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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。 相似文献
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目的 了解蚌埠地区乙型肝炎患者的病毒(HBV)基因型分布情况,并探讨其临床意义。方法 选择在我院就诊的乙型肝炎患者268例,采用荧光定量PCR测定乙肝患者的病毒载量并进行基因分型,同时分析基因型与患者年龄及HBeAg的关系。结果 268份血清标本中有202株为C基因型,占75.37%(202/268),46株为B基因型,20株为B+C型;C基因型的病毒载量和HBeAg阳性率明显高于其他基因型,B基因型感染者的年龄相对较轻。结论 蚌埠地区的乙型肝炎患者基因型以C型为主,B基因型次之,基因型与乙肝患者HBeAg阳性率、年龄等临床指标有一定的相关性。 相似文献
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深圳地区乙型肝炎病毒DNA基因分型与临床的关系 总被引:8,自引:0,他引:8
目的检测血清中乙型肝炎病毒(HBV)DNA并进行基因分型。方法采用聚合酶链反应(PCR)扩增HBV DNA后,用不同的探针,利用微板杂交法将其分为6个亚型。结果150例HBV DNA阳性病人中,B型50例、C型36例.D型13例、F型3例和A型1例,没有发现E型。另有47例为混合型,其中以B、D型(18例)、C、D型(20例)为主,分别占混合型的38.30%与42.6%。受检病人中,B型感染者丙氨酸氨基转移酶水平明显增高;B型感染者比C型感染者抗HBe阳性率高,分别为64.0%和30.5%,而C型感染者比B型感染者乙型肝炎e抗原阳性率高,分别为47.2%和22.0%;年龄、性别与基因型关系不明显。结论深圳地区HBV基因亚型以B型为主,C型次之,其他较少。基因型与肝炎程度有一定关系。 相似文献
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Fuminaka Sugauchi Etsuro Orito Tomoyoshi Ohno Yasuhito Tanaka Jong-Hon Kang Tomoyuki Kuramitsu Eiji Tanaka Takafumi Ichida Kazuaki Inoue Shiro Iino Shinichi Kakumu Takeshi Okanoue Yoshikazu Murawaki Morikazu Onji Michio Sata Ryuzo Ueda 《Hepatology research》2006,36(2):107-114
Genotypes of hepatitis B virus (HBV) were determined in 485 patients with acute hepatitis B from all over Japan. They were A in 92 (19%), Ba in 26 (5%), Bj in 32 (7%), C in 330 (68%) and D in 5 (1%). Sexual contacts were the main route of transmission in them. Overall, HBV persisted in only 5 of the 464 (1%) followed patients. Genotypes C accounted for more than 68% in northern as well as southern areas, contrasting with genotype A accounting for 34% in and around the Metropolitan areas. During 24 years from 1982 to 2005, genotype A increased from 5% to 33%, while genotype B gradually decreased from 26% to 8%. Fulminant hepatitis was significantly more frequent in infection with genotype Bj (41%) than those with the other genotypes (p < 0.01). The core-promoter double mutation (T1762/A1764) and precore stop-codon mutation (A1896) were more frequent in patients with fulminant than acute self-limited hepatitis (57% versus 15% and 58% versus 10%, respectively, p < 0.01 for both). In conclusion, genotype A distributes unevenly over Japan, prevails in younger patients through sexual transmission and has increased with years. Furthermore, fulminant outcome was more frequent in patients with genotype Bj than those with the other genotypes. 相似文献
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目的了解成人急性乙型肝炎患者流行的病毒基因型及基因变异情况。方法在2010年12月~2014年1月北京佑安医院收治39例急性乙型肝炎患者,采用直接基因测序法和平行等位基因特异性检测技术检测HBV基因型、P区、S区、前C区(PreC)和基本核心启动子(BCP) 区序列。结果在39例患者中,成功测序35例。35例急性乙型肝炎患者感染HBV B基因型12例(36.4%),C基因型21例(57.6%,D基因型2例(6.0%);通过直接测序法和PASS法均检测到同1例患者存在A181S耐药变异,变异病毒占准种池比例达100%;直接测序法检测到1例患者存在S区S132F和W172C变异;1例患者存在PreC/BCP 区G1896A和T1758C变异。结论急性乙型肝炎患者感染病毒基因型与文献报道的慢性乙型肝炎患者感染病毒基因型分布一致,以C型和B型为主,存在P区、S区和PreC/BCP区变异可能,未见不同病毒株感染引起转归的不同。 相似文献
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Distribution of hepatitis B virus genotypes in patients with chronic hepatitis B in Turkey 总被引:5,自引:0,他引:5
AIM: Hepatitis B virus (HBV) strains isolated worldwide has been classified into eight genomic groups deduced from genome comparisons and designated as genotypes A to H. We aimed to investigate prevalence of HBV genotypes and subtypes in Turkey. METHODS: A total of 88 chronic hepatitis B (CHB) patients from 15 hospitals throughout the country were included. Patients who were HBsAg positive in serum at least for 6 mo, who had HBV-DNA in serum and elevation of ALT levels more than two times upper limit of normal, and who had percutaneous liver biopsy within 6 mo were included. Genotyping of HBV was done by restriction fragment length polymorphism (RFLP). The patients received subcutaneous 9 MU interferon-α 2a thrice a week for a period of 6 mo. RESULTS: Genotype D was detected in 78 of 88 (88.7%) patients, however, genotyping failed in two patients (2.3%), while no product was obtained in eight (9.0%) patients. Regarding subtypes, D2 was more prevalent (67 patients between 78% and 85.9%) followed by subtype D2+deletion (seven patients of 78 or 8.9%), subtype D1(three patients of 78% or 3.9%) and subtype D3 (one patient of 78% or 1.3%). Thirty-three patients (37.5%) were HBeAg positive compared to 55 (62.5%) anti-HBe positive patients. The endpoint for the viral response of HBeAg positive patients was 27.2%, while it was found 52.7% in HBeAg negative patients (P<0.05). Long-term persistent viral response was 29.5% for all patients. CONCLUSION: This multi-center study indicates that the predominant genotype with CHB patients in Turkey like in other Mediterranean countries is genotype D. 相似文献
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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HBV DNA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HBV DNA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HBV DNA水平差异无统计学意义(6.37±1.62lg copies/ml对6.29±1.76lg copies/ml),但均高于B和C基因型混合感染患者(5.25±1.65lg copies/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。 相似文献
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Filiz Türe Ozdemir Deniz Duman Deniz Ertem Erol Av?ar Fatih Eren Osman Ozdo?an Cem Kalayci Nuray Aslan Ali Mithat Bozdayi Nurdan T?zün 《The Turkish journal of gastroenterology》2005,16(4):183-187
BACKGROUND/AIMS: There are significant variations in the geographic distribution of hepatitis B virus genotypes throughout the world, and some genotypes are associated with different clinical outcomes. Eight genotypes of human hepatitis B virus (designated A-H) have been described to date. To determine the hepatitis B virus genotypes in Turkish patients with chronic liver disease and compare the results with clinical characteristics of the patients. METHODS: Fifty-four (pediatric: n=25 and adult: n=29) patients with chronic hepatitis B virus infection and with an hepatitis B virus DNA level above 5 pg/ml were entered into the trial. Restriction fragment length polymorphism method was used to determine hepatitis B virus genotype and their restriction fragment length polymorphism patterns. Hepatitis B virus DNA samples of 13 patients were sequenced automatically for further confirmation of restriction fragment length polymorphism results. RESULTS: Genotype D was the dominant genotype in all of our cases. Among six restriction fragment length polymorphism patterns of genotype D reported in the literature, three (D1, D2, D6) were present in our series and D2 was the most frequent restriction fragment length polymorphism pattern (81.5%). No significant differences were observed among different genotype D restriction fragment length polymorphism patterns with respect to patients' serum ALT, AST, and hepatitis B virus DNA titer, but D2 restriction fragment length polymorphism pattern was significantly more common in younger adults compared to D1 restriction fragment length polymorphism pattern. CONCLUSION: Genotype D with D2 restriction fragment length polymorphism pattern is the dominant hepatitis B virus genotype in all age groups in Turkey. 相似文献
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目的:研究HBV家族聚集性感染者的基因型和基因变异特征.方法:选择HBV聚集性感染家族成员内部基因型完全相同的共27个家族84人,其中HBV 携带者(ASC)29例;慢性活动性肝炎(CAH)23 例;肝硬化(LC)21例;肝细胞癌(HCC)11例;用基因芯片法检验前C1896和BCP1762,1764双突变;用测序法检验HBV基因型,并设相应基因型的HBV感染对照组.结果:B型HBV家族聚集感染者ASC率显著大于C型家族(X2=5.0734,P=0.0243,P<0.05); 其BCP1762.1764双突变的发生率也显著低于 C基因型(X2=22.4000,P=0.0000,P<0.001).两种基因型的家族聚集感染者组与相应的对照组相比在疾病临床类型方面均无显著差异.结论:检验HBV家族聚集感染者的基因型和/或BCP区1762、1764基因的双突变可以协助疾病的预测分析. 相似文献
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Pérez-Olmeda M Núñez M García-Samaniego J Ríos P González-Lahoz J Soriano V 《AIDS research and human retroviruses》2003,19(8):657-659
Hepatitis B virus (HBV) genotypes were examined in HIV-infected patients with chronic HBV infection seen in our clinic during 2002. A total of 28 of 1100 individuals (prevalence 2.5%) were found to be HBsAg and HBV-DNA positive. HBV genotypes could be determined in 23 of them. HBV-A was the most common (57%), followed by HBV-D (39%). HBV-A predominated among homosexual men (67%) while HBV-D predominated among intravenous drug users (67%). The presence of serum HBeAg was significantly associated with the HBV-A genotype. 相似文献
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Hepatitis B genotypes correlate with clinical outcomes in patients with chronic hepatitis B 总被引:149,自引:0,他引:149
BACKGROUND & AIMS: Six genotypes (A-F) of hepatitis B virus (HBV) have been identified; however, the genotype-related differences in the pathogenicity of HBV remain unknown. Therefore, we investigated the prevalence of HBV genotypes in Taiwan and the association between distinct genotypes and severity of liver disease in a cross-sectional study. METHODS: Using a molecular method, HBV genotypes were determined in 100 asymptomatic carriers and in 170 patients with histologically verified chronic liver disease and hepatocellular carcinoma (HCC). RESULTS: All genotypes except genotype E were identified in Taiwan, and genotypes B and C were predominant. Genotype C was prevalent in patients with cirrhosis and in those with HCC who were older than 50 years compared with age-matched asymptomatic carriers (60% vs. 23%, P < 0.001, and 41% vs. 15%, P = 0.005, respectively). Genotype B was significantly more common in patients with HCC aged less than 50 years compared with age-matched asymptomatic carriers (80% vs. 52%, P = 0.03). This predominance was more marked in younger patients with HCC (90% in those aged =35 years), most of whom did not have cirrhosis. CONCLUSIONS: Our data suggest that HBV genotype C is associated with more severe liver disease and genotype B may be associated with the development of HCC in young Taiwanese. However, additional large-scale longitudinal studies are needed to confirm the relationship of HBV genotypes to liver disease severity and clinical outcomes. 相似文献