新疆地区居民乙肝病毒基因型的初步研究

目的 初步确定新疆地区乙肝患者乙肝病毒基因型的基本情况.方法 对在新疆地区采集的2000余份血清中检测得到200份HBsAg阳性的血清,利用巢氏PCR法扩增其S基因并测序,用MEGA3软件将测序结果与GenBank中的HBV标准序列进行分析,构建并绘制系统发生树,分析基因型.结果 在200份样本中.127例(63.5%)成功检出基因型,其中B基因型10例(7.8%),C基因型58例(45.7%),D基因型59例(46.5%).结论 新疆地区HBV基因型以C、D型为主,少量为B型.     

广州地区小儿无症状乙肝病毒携带者的基因型研究

目的扩增HBV DNA S基因,建立HBV DNA的分型方法,研究广州地区小儿无症状乙肝病毒携带者(AsC)的基因型. 方法利用PCR-RFLP基因分型方法,扩增HBV DNA S基因,以限制性内切酶AvaII、MboI酶切分型. 结果 60份AsC血清样本,B型37份(62.7%),C型20份(33.3%),B、C混合型2份(3.3%),未分型1份(1.7%),经序列分析证明为C型,C型共35%. 结论 PCR-RELP分型方法经济实用,分型率达98.3%,广州地区小儿乙肝病毒的基因型以B型和C型为主,其中携带者中B型占优势.     

河南省漯河地区乙型肝炎病毒基因型别的初步研究

目的 初步确定流行于河南漯河地区的乙肝病毒的基因型的基本情况.方法 采集河南漯河地区乙型肝炎表面抗原阳性的血清,利用PCR扩增得到HBV的S基因;用DANSTAR软件对其进行核苷酸序列分析,构建系统发生树,分析基因型.结果约75.7%样本的HBV S基因序列位于HBV系统发生树的基因型C,约20%的样本其S基因位于系统发生树的基因型B,近4.3%的样本其S基因位于系统发生树的基因型D.结论 流行于河南漯河地区的乙肝病毒多数为基因型C,少数为基因型B,极少数为基因型D.     

绍兴地区HBV母婴传播的基因型分布与特征

目的了解绍兴地区乙型肝炎病毒（HBV）感染孕妇及其新生儿HBV基因型分布以及母婴传播特征。方法收集HBsAg阳性分娩孕妇及其新生儿外周血,用聚合酶联反应-限制性片段长度多态性（PCR-RFLP）分型法鉴定孕妇及新生儿的HBV基因型,对于PCR-RFLP法不能分型的标本,另进行PCR产物直接测序鉴定HBV基因型。结果993例HBsAg阳性孕妇所生新生儿经联合主动、被动免疫,生产HBsAg阳性新生儿116例,新生儿预防接种的失败率为11.68%。116对母婴所测的HBV基因型中,81对为基因B型（69.83%）,其中1例为混合型（Ba与Bj）,35对为基因C型（30.17%）。新生儿HBV基因型与母亲基因型均一致。结论绍兴地区HBV感染孕妇及其受感染新生儿的基因型分布全部是B型与C型,以B型为主,未见其他基因型,HBV基因型在母婴传播中未见改变。     

山西省乙型肝炎病毒基因型别的初步研究

目的 初步确定流行于山西地区的乙肝病毒的基因型的基本情况。方法 采集山西地区乙型肝炎表面抗原阳性的血清，利用PCR扩增得到HBV的S基因和C基因；用MEGA3软件对其进行核苷酸序列分析，构建系统发生树，分析基因型。结果 约93％样本的HBVS基因和C基因序列均位于HBV系统发生树的基因型C，近7％的样本其S基因和C基因序列位于系统发生树的基因型B。结论 流行于山西地区的乙肝病毒多数为基因型C，少数为基因型B，未发现基因重组现象。     

中国南北两城市乙型肝炎病毒基因型与血清型的构成差异

目的对我国南北两城市530份乙型肝炎(乙肝)病毒(HBV)进行血清型和基因型分型,以了解HBV基因型和血清型分布的特点和差异。方法 对黑龙江省哈尔滨市和广东省廉江市530份乙肝病毒表面抗原(HBsAg)阳性血清进行HBV DNA基因扩增,并对扩增产物直接测序,分析HBV血清亚型和基因型。结果 哈尔滨和廉江市HBV血清型以adrq为最多,分别为87.2％和73.5％,其次为adw2,分别为12.0％和25.7％,其分布差异有非常显著意义(P<0.001);两市HBV基因型均以C型为主,分别为87.8％,73.2％,其次为B型,分别为12.2％和26.1％,其分布差异有非常显著意义(P<0.001),在廉江市仅有1例D型,1例B、C混合型。结论我国南北两城市HBV血清亚型和基因型的构成较为单纯,均只有两个类型,其比例构成差异有非常显著意义。     

应用多重巢式PCR法快速鉴定广西HIV-1主要亚型

目的 建立快速鉴定广西HIV-1主要流行亚型CRF01_AE、CRF07_BC、CRF08_BC、B和C的多重巢式PCR方法.方法 针对HIV-1 gag基因设计CRF01_AE、CRF07_BC、CRF08_BC、B和C亚型特异性引物,建立多重巢式PCR法,用该法鉴定来自广西的HIV-1毒株的亚型,并与基因测序法的鉴定结果比较.结果 多重巢式PCR法能正确鉴别5种已知亚型的样本,10份HIV阴性样本均无扩增,在72份未知亚型样本中,正确鉴定出66份,分别属于CRF01_AE、CRF07_BC、CRF08_BC、B亚型,多重巢式PCR法的灵敏度为91.7%,特异度达100%.结论 多重巢式PCR法能准确鉴定广西CRF01_AE、CRF07_BC、CRF08_BC和B亚型毒株,是一种简便、快速、低成本的亚型鉴定方法.     

两种型特异性引物聚合酶链反应法检测乙型肝炎病毒基因型和亚型的评价

目的 比较并评价本室创建的方法和日本Naito 等建立的乙型肝炎病毒(HBV)型特异性引物聚合酶链反应(PCR)基因分型法.方法 分别用新方法和Naito法检测系列稀释的含有A、D基因型和B1、C2亚型HBV基因组的质粒及不同浓度比例的B1/C2亚型混合质粒,评价两种方法的灵敏度和特异度.此外,用该两种方法分别对深圳市、河北邯郸市、新疆乌鲁木齐市送检的113份HBV DNA阳性慢性乙型肝炎患者的血清标本进行HBV基因分型,并对该两种分型方法检测结果不一致的血清样本,用PCR产物直接测序法分析.结果 两种方法对单一型别的质粒样本灵敏度无差异,但新方法对B/C混合型质粒样本的灵敏度及对各型别质粒样本的特异度明显优于Naito法.该两种方法检测单一基因型血清标本的灵敏度无差异,但新方法对B/C混合型血清样本的检出率更高.该两种方法检测结果总符合率为83.2%(94/113),不一致率为16.8%(19/113).从两法分型不一致的19份血清标本中,选取15份以PCR产物直接测序法进行分析,结果与新方法检测结果完全一致,而与Naito法不一致.结论 本室建立的HBV型特异性引物-PCR基因分犁法具有较高灵敏度,其特异度也明显优于目前临床上常用的Naito法,适用于HBV 基因型和基因亚型的临床及流行病学研究.     

ABO基因278C>T突变导致Bw12一例

目的 对1例ABO疑难血型样本进行血清型和基因型鉴定.方法 用血清学方法对标本进行ABO血清型检测,根据血清型结果,选取ABO基因亚型检测试剂盒,用聚合酶链反应-序列特异性引物法进行基因亚型鉴定,用直接测序法对ABO基因的第6、7外显子进行序列测定.结果 血清学结果显示,该样本的红细胞上具有高凝集强度的A抗原和中等凝集强度的B抗原,样本血浆中含有弱凝集强度的抗-B抗体,初步判定为ABw型;B亚型基因分型试剂的结果显示,该样本ABO基因为ABw12型;直接测序结果显示,该标本ABO基因第6外显子序列为278CT、297GA,第7外显子序列为467CT、526CG、657CT、703GA、796CA、803GC、930GA杂合,即在基因型A102/B101的基础上,该序列nt278位发生了C＞T杂合突变,经与血型抗原基因变异资料库的数据比对,确定突变的等位基因为Bw12,基因型为A102/Bw12.结论 中国人群中发现A102/Bw12基因型.     

广州地区小儿无症状乙肝病毒携带者的基因型研究

目的：扩增HBV DNA S基因，建立HBV DNA的分型方法，研究广州地区小儿无症状乙肝病毒携带者（AsC）的基因型。方法：利用PCR－RFLP基因分型方法，扩增HBV DNA S基因，以限制性内切酶AvaⅡ、MboI酶切分型。结果：60份AsC血清样本，B型37份（62．7％），C型20份（33．3％），B、C混合型2份（3．3％），未分型1份（1．7％），经序列分析证明为C型，C型共35％。结论：PCR－RELP分型方法经济实用，分型率达98．3％，广州地区小儿乙肝病毒的基因型以B型和C型为主，其中携带者中B型占优势。     

Hepatitis B virus subgenotype C2 is the most prevalent subgenotype in northeast China

The geographical distribution of hepatitis B virus (HBV) subgenotypes and their clinical implications in patients with acute and chronic hepatitis B in the Heilung-kiang province of northeast China were investigated. Nested PCR and multiplex PCR were performed with genotype-specific primers and with subgenotype-specific primers to identify genotypes and subgenotypes from serum samples of 412 HBV infections including 69 with acute self-limited hepatitis (ASH) and 343 with chronic hepatitis (CH). A total of 361 samples were genotyped and 304 were further subgenotyped. The most common HBV genotype was C (93.63%, 338/361), with subgenotype group C2 (83.73%, 283/338) predominating. Genotype B was also found and subgenotype B2 predominated within this genotype. Out of 69 infected patients with ASH, 48 were identified as genotype C and all belonged to subgenotype C2. Of 343 infected patients with CH, 313 were genotyped and 256 were subgenotyped; amongst these, C2 (91.80%, 235/256), B2 (7.42%, 19/256) and mixed subgenotypes B2 and C2 (0.78%, 2/256) were found. In HBV subgenotype C2 infections, ASH had a higher ratio of women than CH patients. These results show that HBV subgenotypes C2 and B2 were found in Heilung-kiang province of northeast China. In ASH and CH groups, the distributions of subgenotypes were coincident with C2, the predominant subgenotype. Analysis of the association between subgenotype and the outcomes of HBV infection was inconclusive in our study.     

Genetic study of hepatitis B virus in Indonesia reveals a new subgenotype of genotype B in east Nusa Tenggara

The hepatitis B virus (HBV) genotype is associated with viral anthropological history, clinical outcome of disease and response to treatment. This study examines the HBV genotypes in Indonesia. HBV genotypes were determined by whole-genome sequencing and from the sequence of the Pre-S2 and S regions in a larger series. Two HBV genotypes, B (HBV/B) and C (HBV/C), were predominant. Three previously reported HBV/B subgenotypes were identified, with certain population association: HBV/B2 (HBV/Ba) was found mostly in Indonesians of Chinese ethnic origin, HBV/B3 was dominant among the Javanese, and HBV/B5, reported earlier from the Philippines, was also discovered, albeit at low frequency. Two other subgenotypes, HBV/B4 from Vietnam and HBV/B6, recently reported from the Arctic region, were not found. A novel subgenotype, HBV/B7, was recognized, associated with populations of the Nusa Tenggara islands in eastern Indonesia. Characteristic differences in HBsAg serotype and single nucleotide polymorphisms (SNPs) in the Pre-S2 region distinguish HBV/B7 from other HBV/B subgenotypes and further establish the new HBV subgenotype.     

Virological and clinical characteristics on reactivation of occult hepatitis B in patients with hematological malignancy

The virological characteristics of hepatitis B virus (HBV) implicated in the reactivation of occult hepatitis B in patients who have received hematopoietic stem-cell transplantation or chemotherapy for the hematological malignancy are not well defined. Twenty-eight HBsAg-negative patients who received hematopoietic stem-cell transplantation and 138 HBsAg-negative patients treated for malignant lymphoma with chemotherapy including rituximab were enrolled. Three of the 28 patients (10.7%) received hematopoietic stem-cell transplantation and one of the 138 (0.72%) patients treated for malignant lymphoma with chemotherapy developed de novo HBV hepatitis. Anti-HBc was detected in four and anti-HBs in two patients. Genotype Bj was detected in two and C in two of they all possessed wild-type sequences in the core promoter region. A precore stop mutation (A1896) was detected in a patient with genotype Bj who developed fulminant hepatic failure. HBV DNA was detected in pretreatment HBsAg-negative samples in two of four patients, and the HBV genome sequence identified from sera before chemotherapy and at the time of de novo HBV hepatitis showed 100% homology. In an in vitro replication model, genotype Bj with the A1896 clone obtained from a fulminant case had a replication level much higher than clones obtained from de novo hepatitis B patients with genotype Bj or C with G1896. In conclusion, this is the first report demonstrating de novo hepatitis B from the reactivation of occult HBV infection confirmed by molecular evolutional analysis. The fulminant outcome of HBV reactivation can be associated with genotype Bj exhibiting high replication due to the A1896 mutation.     

Five subgenotypes of hepatitis B virus genotype B with distinct geographic and virological characteristics

Several hepatitis B virus (HBV) subgenotypes, HBV/A1, A2, Bj and Ba, have been reported with respect to clinical differences among patients infected with these subgenotypes. The population genetics and phylogeography of HBV were investigated based on the complete genome sequences of 484 isolates with 108 from our chronic hepatitis B patients and the remaining from the GenBank database. Besides genotypes A-H (HBV/A-H), five subgenotypes were identified among 169 HBV/B isolates by phylogenetic analysis and nucleotide divergence. There were 27 isolates of subgenotype B(1) (HBV/B(1)) restricted to Japan, 104 isolates of HBV/B(2) with the widest distribution in most Asian countries, 4 isolates of HBV/B(3) restricted to Indonesia, 32 isolates of HBV/B(4) restricted to Vietnam, and 7 isolates of HBV/B(5) restricted to Philippines. HBV/B(2)-B(5) isolates carried a recombination with HBV/C over the precore and core genes. In addition to the characteristics of HBV/B(1)-B(5) at some cis-acting elements, the precore stop-codon mutant (G1896A) was significantly different among HBV/B(1), HBV/B(2), and HBV/B(4) (70.3%, 31.7%, 53.0%, P=0.001), while no such mutation was found in HBV/B(3) and B(5). Among characteristics of the HBV/B(1)-B(5) amino acid sequences, serotype adw (K(122)) was exclusive among HBV/B(1), HBV/B(2), and HB V/B(3) isolates, while serotype ayw (R(122)) was among the HBV/B(4) and HBV/B(5) isolates. Furthermore, distinct variations of T cell and B cell recognition epitopes within surface and core proteins were also found among these subgenotypes. In conclusion, subgenotypes HBV/B(1)-B(5) exhibited distinct geographical distributions, virologic characteristics, and probable clinical implications.     

Comparison of hepatitis B virus subgenotypes in patients with acute and chronic hepatitis B and absence of lamivudine-resistant strains in acute hepatitis B in Japan

Hepatitis B virus (HBV) has been classified into eight genotypes and can be further divided into several subgenotypes that have different geographic distributions. Because of increased human migration, the prevalence of rare subgenotypes is increasing in Japanese patients with acute hepatitis B. Lamivudine-resistant strains of HBV have begun to emerge in association with chronic hepatitis B. The aim of this study was to investigate the distribution of HBV subgenotypes and lamivudine-resistant strains in patients in Japan with acute hepatitis B. One hundred twenty-three patients with acute hepatitis B and 123 with chronic hepatitis B were studied. HBV subgenotypes and lamivudine-resistance mutations were determined by direct sequencing of the preS and polymerase region, respectively. HBV subgenotypes Aa (n=3), Ae (n=23), Ba (n=7), Bj (n=3), Cs (n=7), Ce (n=76), D (n=2), and H (n=2) were detected in patients with acute hepatitis. In patients with chronic hepatitis, HBV subgenotypes Ae (n=4), Ba (n=1), Bj (n=18), and Ce (n=100) were found. Non-common Japanese subgenotypes, that is, non-Bj and non-Ce, were detected more frequently in patients with acute hepatitis (35.8%) than in patients with chronic hepatitis (4.1%) (Odds ratio, 0.076; 95%CI, 0.029-0.200; P<0.0001). Lamivudine-resistance mutations were detected in chronic hepatitis patients with breakthrough hepatitis but not in other patients. In conclusion, the prevalence of uncommon Japanese HBV subgenotypes is expected to increase, although lamivudine-resistant strains have not yet been found in patients with acute hepatitis B.     

Virological features of hepatitis B virus-associated nephropathy in Japan

Hepatitis B virus (HBV)-associated nephropathy is considered as an immune-mediated disorder which is dependent on interactions between viral, host, and environmental factors. But there are few reports that investigated the relationship between the development of HBV-associated nephropathy and HBV genotypes and the mutations. To clarify the relationship between nephropathy and HBV genotype in Japan, six male patients with HBV-associated nephropathy were examined. The complete genome sequences of HBV were determined directly and the specific mutations associated with the development of HBV-associated nephropathy were examined by comparison of the alignments along with consensus sequences [HBV/A1 (Aa), A2 (Ae), B1 (Bj), B2 (Ba), C1 (Cs) and C2 (Ce)] retrieved from international database. The mean age of the six patients was 33.5 years. HBeAg was found in all patients and serum HBV-DNA levels were relatively high. Histological findings of renal tissues indicated five cases of membranous nephropathy and one membranoproliferative glomerulonephritis. HBV genotypes from the six patients were two HBV/A1, two A2 and two C2, suggesting HBV/A was predominant. G1862T mutation was observed in the two HBV/A1 patients, resulting in the pre-core amino acid substitution with a switch from valine (Val) to phenylalanine (Phe). Only one patient had core deletions. It is concluded that HBV/A may be associated with membranous nephropathy, but little relationship between HBV gene mutations and the development of HBV-associated nephropathy was observed.     

乙型肝炎病毒C启动子区C1673T/C1799G联合突变的意义

目的 探讨HBV(乙型肝炎病毒)C启动子区(CP) C1673T/C1799G联合变异的生物学和临床意义.方法 136名慢性HBV感染者,包括无症状携带者(ASC) 25例,慢性乙型肝炎(CHB)38例,慢性重型乙型肝炎(CSHB) 24例,乙肝肝硬化(LC) 36例,肝细胞肝癌(HCC)13例,用半巢氏PCR的方法结合直接测序法检测HBV基因亚型及CP区变异,分析C1673T/C1799G联合变异在不同基因亚型中的发生率及与HBV复制、e抗原表达和不同慢性HBV感染疾病谱的关系.结果 本组病例中,Ba亚型110例,Bj亚型1例,C1亚型7例,C2亚型8例,C1673T/C1799G联合变异发生率为80.9％,其中在Ba亚型中为96.4％,C1亚型为14.3％,C2亚型为12.5％,Ba亚型与C1和C2亚型比较,差异有统计学意义(P ＜0.0001).变异组HBV DNA载量与非变异组比较差异无统计学意义(P＞0.05);e抗原阳性组变异率为71.4％,e抗原阴性组为87.5％,两组比较差异有统计学意义(P＜0.05).从ASC、CHB、CSHB、LC到HCC组,变异的发生率差异无统计学意义(P＞0.05).结论 C1673T/C17991G联合变异常见于Ba基因亚型,该变异不影响HBV DNA的复制水平,可能与不同慢性HBV感染疾病谱无关.     

Clinical and virological characteristics of hepatitis B virus subgenotypes Ba, C1, and C2 in China

Hepatitis B virus (HBV) subgenotypes Ba, C1 (Cs), and C2 (Ce) are the most prevalent HBV variants in China. To investigate the virological characteristics of these subgenotypes and their clinical implications, we enrolled a cohort of 211 patients in the Guangdong Province of China, including 132 with chronic hepatitis B virus infection (CH), 32 with liver cirrhosis (LC), and 47 with hepatocellular carcinoma (HCC) according to clinical examination, liver function test, and ultrasonograph results. Overall, HBV Ba was found in 51.2% (108/211), HBV C1 in 33.6% (71/211), and HBV C2 in 15.2% (32/211) of the cases. The distribution of HBV genotype C was greater among patients in the LC and HCC groups than among patients in the CH group, while the distribution of HBV genotype B was greater among the CH patients than among the LC and HCC patients. No significant differences in clinical features were found among patients with HBV Ba, C1, and C2. Virologically, HBV C1 had the strongest association with the A1762T G1764A double mutation, while the mutation at position 1896 resulting in A (1896A) was uncommon. In contrast, HBV Ba had the highest frequency of 1896A but the lowest of A1762T G1764A, and HBV C2 had intermediate frequencies of these mutations. Mutations of 1653T and 1753V were specifically associated with HBV C2 and C1, respectively. Multivariate analyses showed that the 1653T, 1753V, and A1762T G1764A mutations and patient age significantly increased the risk of HCC development. In conclusion, HBV Ba, C1, and C2 have different mutation patterns in the enhancer II/core promoter/precore region. Therefore, genotyping and detecting the 1653T and 1753V mutations, in addition to the A1762T G1764A double mutation, might have important clinical implications as predictive risk factors for hepatocarcinogenesis.     

Genotype X/C recombinant (putative genotype I) of hepatitis B virus is rare in Hanoi,Vietnam—genotypes B4 and C1 predominate

There are eight known genotypes of hepatitis B virus, A–H, and several subgenotypes, with rather well‐defined geographic distributions. HBV genotypes were evaluated in 153 serum samples from Hanoi, Vietnam. Of the 87 samples that could be genotyped, genotype B was found in 67 (77%) and genotype C in 19 (22%). All genotype C strains were of subgenotype C1, and the majority of genotype B strains were B4, while a few were B2. The genotype X/C recombinant strain, identified previously in Swedish patients of indigenous Vietnamese origin, was found in one sample. This variant, proposed to be classified as genotype I, has been found recently also by others in Vietnam and Laos. The current study indicates that the genotype X/C recombinant may represent approximately 1% of the HBV strains circulating in Vietnam. J. Med. Virol. 82:1327–1333, 2010. © 2010 Wiley‐Liss, Inc.