我国九个地区丁型肝炎病毒感染状况及其与乙型肝炎…

对１５４５例各类乙型肝炎病毒表面抗原（ＨＢｓＡｇ）阳性肝病和无症状ＨｓＡｇ携带者的血清进行了乙型肘炎病毒（ＨＢＶ）与丁型肝炎病毒（ＨＤＶ）感染标记物的测定。结果表明，ＨＤＶ感染率为１３．０１％，其中ＨＤＡｇ和抗－ＨＤ阳性率分别为２．９１％和１０．０９％。而且在全国九个地区均有ＨＤＶ感染者存在，说明其分布是较为广泛的。同时还表现出，男性高于女性，慢性肝炎、重型肝炎及原发性肝癌高于急性肝炎和无症状ＨＢ     

我国九个地区丁型肝炎病毒感染状况及其与乙型肝炎病毒复制指标关系的研究

对１５４５例各类乙型肝炎病毒表面抗原（ＨＢｓＡｇ）阳性肝病和无症状ＨＢｓＡｇ携带者的血清进行了乙型肝炎病毒（ＨＢＶ）与丁型肝炎病毒（ＨＤＶ）感染标记物的测定。结果表明，ＨＤＶ感染率为１３．０１％，其中ＨＤＡｇ和抗－ＨＤ阳性率分别为２．９１％和１０．０９％。而且在全国九个地区均有ＨＤＶ感染者存在，说明其分布是较为广泛的。同时还表现出，男性高于女性，慢性肝炎、重型肝炎及原发性肝癌高于急性肝炎和无症状ＨＢｓＡｇ携带者。提示ＨＢＶ与ＨＤＶ合并感染或重叠感染可能导致病情加重和感染的慢性化。本项研究结果还揭示，在ＨＢＶ与ＨＤＶ合并或重叠感染时，可能对ＨＢＶ的复制指标（ＨＢｅＡｇ·ＨＢＶＤＮＡ）有一定的抑制现象。     

丁型肝炎病毒在乙肝病毒感染者中分布状况及其分子生物学研究

通过对丁型肝炎病毒（ＨＤＶ）的放免法检测和巢式ＰＣＲ技术检测，在１２０例乙肝病毒感染者中，检测到１３例ＨＤＶ标记阳性（阳性率为１０．８％），５０例乙肝病毒携带者，３例抗ＨＤＶ－ＩｇＭ阳性。经对照所得乙肝患者同乙肝病毒携带者的ＨＤＶ阳性差异结果支持ＨＤＶ的存在常伴随着谷丙转氨酶（ＡＬＴ）活性上升的观点。实验结果还提示了ＨＤＶ和乙型肝炎病毒（ＨＢＶ）重叠感染易发生重症或慢性肝炎。地ＨＤＶ的检测方法进行     

成年树鼩实验感染丁型肝炎病毒的初步研究

在证实成年树可感染人乙型肝炎病毒（ＨＢＶ）的基础上，进行了丁型肝炎病毒－乙型肝炎病毒（ＨＤＶ／ＨＢＶ）实验感染的探索。ＨＤＶ／ＨＢＶ阳性人血清经同时和重叠感染方式接种于成年树后，定期留取感染树血清及肝组织，检测血清中ＨＢｓＡｇ、ＨＤＡｇ、抗－ＨＤ、ＨＢＶＤＮＡ及ＨＤＶＲＮＡ，初步探讨成年树实验感染ＨＤＶ的可能性。研究发现：①成年树对人ＨＢＶ易感，ＨＢｓＡｇ阳性率为７５％，其血清学反应及肝组织病理改变与文献报道一致；②ＨＤＶ可通过同时和重叠两种方式感染成年树，感染树血清中可出现ＨＤＡｇ及抗－ＨＤ，经分子杂交证实其血清及肝内均有ＨＤＶＲＮＡ和ＨＢＶＤＮＡ存在，且可导致明显的肝细胞损伤。结果提示：成年树既可感染人ＨＢＶ也可感染ＨＤＶ，可作为研究ＨＤＶ人工感染的实验动物；成年树感染ＨＤＶ后的特点与人及黑猩猩感染时十分相似，能较好地反映人丁型肝炎的真实情况。     

52 例病毒性肝炎患者肝组织中丙型肝炎病毒 RNA 和 HCAg-NS3 的测定

应用地高辛标记探针原位杂交法和单克隆抗ＨＣＶ－ＮＳ３－ＨＲＰ建立直接酶标免疫组化法分别测定５２例肝炎患者肝组织ＨＣＶＲＮＡ和ＨＣＡｇ－ＮＳ３。结果抗ＨＣＶ阳性组ＨＣＶＲＮＡ检出率５７．１％（１６／２８），ＨＣＡｇ－ＮＳ３检出率５３．６％（１５／２８）；抗ＨＣＶ阴性组其两项检出率均为１２．５％（３／２４）。肝组织中ＨＣＶＲＮＡ阳性物呈蓝紫色细小颗粒存在于肝细胞核或胞浆内，其在肝小叶中的分布可分为３型，即弥漫型、局灶型、散在型。肝组织中ＨＣＡｇ－ＮＳ３阳性物呈棕黄色细小颗粒分布于肝细胞核或胞浆内，以单个或数个阳性细胞散布于肝小叶中。２３例ＨＣＶＲＮＡ或／和ＨＣＡｇ－ＮＳ３阳性病例以肝炎后肝硬化（ＬＣ）病例占多数（１４／２３），其次为慢性重型肝炎（ＣＳＨ）和中度慢性肝炎（ＣＡＨ）。此两种检测方法具有较高符合率（９０．４％，４７／５２），表明病毒核酸及其表达产物均存在于肝细胞内，与ＨＣＶ感染密切相关。这为ＨＣＶ感染诊断提供了直接依据，有利于研究ＨＣＶ感染中病毒复制、慢性化进程、抗病毒治疗监测及重叠感染时病毒相互关系。     

检测丁型肝炎病毒RNA打点杂交法的建立及其应用

经缺口平移法以ａ－３２Ｐ－ｄＣＴＰ标记１．０ｋｂ的丁型肝炎病毒（ＨＤＶ）ｃＤＮＡ片段为探针，采用蛋白酶Ｋ直接从血清中提取ＨＤＶＲＮＡ，建立了检测血清中ＨＤＶＲＮＡ的打点杂交法，其灵敏性可达１ｐｇ水平，与乙型肝炎病毒（ＨＢＶ）ＤＮＡ无交叉杂交反应；并应用于检测我国５９４９份ＨＢｓＡｇ阳性血清中的ＨＤＶＲＮＡ，共检出１７６份ＨＤＶＲＮＡ阳性，检出率为２．９５％。     

广西不同人群庚型肝炎病毒感染状况的调查

目的 了解庚型肝炎病毒（ＨＧＶ）感染在广西柳州地区不同人群中的感染状况，并比较静脉毒瘾者与健康体检者ＨＧＶ，乙型肝炎病毒（ＨＢＶ）、丙型肝炎病毒（ＨＣＶ）与ＨＧＶ共同感染的特点，方法 应用酶联免疫法（ＥＬＡ）检测抗－ＨＧＶ，抗－ＨＣＶ，ＨＢＶＭ（ＨＢｓＡｇ，ＨＢｓＡｂ，ＨＢｅＡｇ，ＨＢｅＡｂ，ＨＢｃＡｂ），并对抗－ＨＧＶ阳性者随机抽取２０例（１９％）进一步用逆转录套式ＰＣＲ（ＰＴ－ｎＰＣＲ）法检测     

乙型肝炎病毒在丁型肝炎病毒包装中的作用机理研究

用含ＨＢＶ全基因和ＨＢＶ大、中、小分子表面蛋白基因的真核细胞表达质粒（ＣＭＶ－ＨＢＶ、ＣＭＶ－ＬＳ、ＣＭＶ－ＭＳ、ＣＭＶ－Ｓ）分别与含ＨＤＶ ｃＤＮＡ三聚体的重组质粒共转染ＣＨＯ细胞。转染后３天在上述４种转染细胞内及培养上清中均检出了ＨＤＶ ＲＮＡ和ＨＤＡｇ表明上述４种转染细胞的培养上清中均有ＨＤＶ病毒颗粒的包装和分泌。提示：ＨＤＶ病毒的包装可能仅需ＨＢＶ Ｓ 基因及其小分子表面蛋白的辅助。     

丁型肝炎病人肝组织HDVRNA与 HBVDNA的表达及关系

目的 探讨丁型肝炎病人肝组织中ＨＤＡｇ、ＨＤＶＲＮＡ与ＨＢＶＤＮＡＧ表达及关系。方法 应用免疫组化和原位杂交技术检测了７９全丁型肝炎病人肝组织ＨＤＡｇ、ＨＤＶＲＡＮ～ＨＢＶＤＮＡ表达,以５２例型肝炎病人肝组织作对照。结果 丁型肝炎ＨＢＶＤＮＡ检出率（２７％）低于乙型肝炎（４４％）（Ｐ〈０．０５）。在坏死灶边缘肝细胞和气球样变肝细胞浆内有大量的ＨＤＶＲＮＡ蓄积或ＨＤＡｇ呈强型强表达,ＨＤＶＲＮＡ表达     

血清学标志阴性的非甲～戊型肝炎的病原学研究

目的对血清学标志阴性的非甲～戊型肝炎进行病原学研究。方法用ＨＢＶＰＣＲ、ＨＣＶＲＴ－ＰＣＲ和ＨＥＶＲＴ－ＰＣＲ分别检测血清学标志阴性的非甲～戊型肝炎患者血清，并对其部分阳性产物进行克隆测序。结果８７例非甲～戊型肝炎血清ＨＢＶＤＮＡ均为阴性，９例（１０．３％）为ＨＣＶＲＮＡ阳性，部分经测序证实为ＨＣＶ１ｂ亚型；余７８例为ＨＢＶＤＮＡ和ＨＣＶＲＮＡ均阴性。该７８例中，１４例因无血清未作ＨＥＶＲＮＡ检测，余６４例中４９例（７６．６％）为ＨＥＶＲＮＡ阴性，１５例（２３．４％）为ＨＥＶＲＮＡ阳性。经序列分析显示，其中９例为典型的中国ＨＥＶ株基因序列，６例变异较大，与典型的中国株基因序列的同源性仅为８０％左右。４９例ＨＢＶＤＮＡ、ＨＣＶＲＮＡ和ＨＥＶＲＮＡ均阴性的血清中１６例（３２．６％）ＨＧＶＲＮＡ阳性。由此可见，该８７例中至少有９例为ＨＣＶ感染，１５例为ＨＥＶ感染，１６例为ＨＧＶ感染。结论对血清学标志阴性的非甲～戊型肝炎的病人应该用ＰＣＲ法进行病原学分型，以明确其诊断     

Hepatitis B and D virus infection among drug abusers in Taiwan

Approximately 15 to 20% of the general population in Taiwan are chronic hepatitis B surface antigen (HBsAg) carriers. However, the incidence of hepatitis D virus (HDV) infection is low (5-8%) in patients with HBsAg-positive chronic liver diseases in this area. To evaluate the prevalence of hepatitis B virus (HBV) and HDV infection among drug abusers in Taiwan, serum samples were collected from 152 drug abusers at the Taipei Municipal Anti-Narcotic Institute and test for HBV and HDV markers. Of these, 24 (15.8%) were HBsAg positive, and only 15 (9.9%) were seronegative for all HBV markers. Of the 115 intravenous drug abusers, serum antibody to hepatitis D antigen (anti-HD) was positive in 78.9% of 19 persons who were HBsAg positive, and in 7.5% of 80 persons who were positive for antibody to HBsAg (anti-HBs). Anti-HD was not detected in the sera from all 37 nonintravenous drug abusers regardless of the status of their HBV markers. Also, none of 63 asymptomatic HBsAg carrier pregnant women or 23 patients with acute type B viral hepatitis had measurable anti-HD in their sera. Thus, the high frequency of HDV detected among Chinese HBsAg carrier intravenous drug abusers in Taiwan is similar to that reported in Western countries.     

Hepatitis D virus infection among intravenous drug abusers in Taiwan: analysis of risk factors and liver function tests

To investigate the prevalence of hepatitis D virus (HDV) and hepatitis B virus (HBV) infection among intravenous drug abusers in Taiwan, a total of 761 male prisoners, including 680 intravenous drug abusers, were studied for serological markers of HBV and HDV. Questionnaires were distributed to evaluate the risk factors for HDV infection and also to estimate the strength of association among HDV infection and the risk factors. HBV infection was common, and the positive rates of HBV markers between intravenous drug abusers and non-drug abusers were not statistically different. However, the positive rate of the antibody to HDV was significantly higher among intravenous drug abusers than among non-drug abusers (21.3% vs. 8.6%). Of 131 chronic HBV carriers with intravenous drug abuse, 119 (91%) were anti-HD positive. Using multiple logistic regression models, we found that the most important risk factor for HDV infection was hepatitis B surface antigen (HBsAg) carriage, and intravenous drug addiction the next. A matched case-control study also was conducted to compare liver function tests among both anti-HD- and HBsAg-positive group anti-HD-negative, and HBs-AG-positive group as well as those with neither positive. Statistically significant difference in liver function tests was not found. It is concluded that the HBsAg carriers with intravenous drug abuse in Taiwan are commonly HDV infected with and that the infection does not seem to affect the liver as assessed by liver function tests.     

Diagnostic significance of IgM antibody to hepatitis delta virus in fulminant hepatitis B

The prevalence of hepatitis delta virus (HDV) infection was studied in 25 adult patients with fulminant hepatitis who were admitted consecutively to our unit from February, 1986, to September, 1988. Enzyme and radioimmunoassays were used for the detection of serological markers of HAV, HBV, and HDV (HDAg, IgM anti-HD, total [IgG] anti-HD) infections. Two hundred twenty-nine serum samples (three to 19 samples/patient) were tested for serological markers of HDV infection. Of the 25 patients, 17 (68%) were HBsAg-positive, and the remaining eight (32%) were HBsAg-negative on admission to the hospital. All patients were seropositive for IgM anti-HBc. Serological markers of HDV infection were detected in 13 (52%) of the 25 patients. In particular, HDV infection was observed in nine (53%) of the 17 HBsAg-positive and in four (50%) of the eight HBsAg-negative patients with type B fulminant hepatitis. Survival was 16.7% for patients with hepatitis B and 57.8% for patients with B and D coinfection. Coinfections were responsible for fulminant hepatitis in 100% of drug addicts and 40% in patients who were not drug addicts. All patients with HBV/HDV coinfections became seropositive for IgM anti-HD. The results show that HDV infection has a significant role (52%) in type B fulminant hepatitis in an area with a moderate prevalence of HBV infections, that it should be tested in cases with early clearance of HBsAg, and that it does not seem to be accompanied by a high fatality rate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prevalence and clinical significance of hepatitis D virus co-infection in patients with chronic hepatitis B in Korea

Hepatitis D virus (HDV) infection can cause severe acute and chronic liver disease in patients infected with hepatitis B virus (HBV). Despite the significant decline in the global HDV infection, it remains a major health concern in some countries. This study aimed to investigate the prevalence and clinical features of HDV co-infection in patients with chronic HBV infection in Korea, where HBV infection is endemic. Nine hundred forty patients [median age, 48 (18-94) years; men, 64.5%] infected chronically with HBV were enrolled consecutively. All patients who were positive for hepatitis B surface antigen (HBsAg) for at least 6 months and were tested for anti-HDV. A portion of the HDV delta antigen was amplified, sequenced, and subjected to molecular and phylogenetic analysis using sera from the patients who were anti-HDV positive. Clinical features and virologic markers were evaluated. Inactive HBsAg carriers, chronic hepatitis B, cirrhosis and hepatocellular carcinoma accounted for 29.5%, 44.7%, 17.9%, and 8.0%, respectively. Only three patients were positive for anti-HDV, corresponding to a 0.32% positive rate. All patients who were positive for anti-HDV were inactive HBsAg carriers. HDV RNA could be amplified by PCR from the sera of two patients. Phylogenetic analysis showed that both carried HDV genotype 1. In conclusion, the prevalence of HDV infection is very low (0.32%) in Korea. All HDVs were genotype 1 and detected in inactive HBsAg carriers. Therefore, HDV co-infection may not have a significant clinical impact in Korean patients with chronic HBV infection.     

乙型肝炎病毒感染者血清中丁型肝炎病毒标志物检测分析

目的 了解丁型肝炎病毒(hepatitis delta virus,HDV)感染标志物在乙型肝炎病毒(hepatitis B virus,HBV)感染者中的分布状况并分析其临床意义.方法 收集HBV感染者的临床资料和血清样本,通过酶联免疫分析法检测其血清HBV感染五项指标、HDVAg和Anti-HDV,结合临床诊断和生化指标进行分析.结果 收集HBV感染者样本462例,其中无症状携带者210例,慢性肝炎175例,急性乙肝35例,肝纤维化42例,检出HDV感染率为4.8％,男性显著高于女性,肝纤维化组的HDV感染率最高,为9.5％,其次为慢性肝炎的6.9％,45～60岁人群的HDV感染率为7.8％,显著高于其他年龄段.结论 慢性乙肝和肝纤维化病例的HDV感染显著增高,提示HDV感染与肝病的严重程度相关,建议对肝病患者开展血清HDV感染标志物检查,鉴别是否存在HDV重叠感染.     

Hepatitis B virus concentrations in serum determined by sensitive quantitative assays in patients with established chronic hepatitis delta virus infection.

To clarify the correlation between hepatitis B virus (HBV) DNA levels and serum alanine aminotransferase (ALT) levels in patients with established chronic hepatitis delta virus (HDV) infection, sensitive HBV quantitative assays were used for the study. Thirty-four consecutive patients with chronic liver disease who were positive for both hepatitis B surface antigen (HBsAg) and antibody to HDV (anti-HDV), including 19 patients with chronic hepatitis, 8 patients with liver cirrhosis and 7 patients with hepatocellular carcinoma. All were negative for hepatitis Be antigen (HBeAg) and positive for antibody to HBeAg. HBV DNA was detected in 25 (73.5%) of the 34 patients using real-time detection PCR, and the HBV DNA levels of these patients were significantly lower compared with HBeAg status and ALT level-matched patients with chronic liver disease positive for HBsAg but negative for anti-HDV. There was no correlation between serum HBV DNA and ALT levels among the 34 patients with chronic liver disease positive for anti-HDV. Whereas serum ALT levels in anti-HDV-positive HBsAg carriers with HDV RNA were significantly higher than those without HDV RNA. Liver damage in patients with established chronic HDV infection may be caused mainly by ongoing HDV infection not by HBV replication.     

原位杂交法检测肝组织中丁型和乙型肝炎病毒核酸

利用国外引进的重组质粒获得纯化基因片段，分别以随机引物法和ＰＣＲ法制备地高辛素标记的ＨＢＶＤＮＡ探针和ＨＤＶｃＤＮＡ探针。用原位杂交法检测了石蜡包埋的肝组织切片ＢＶＤＮＡ和ＨＤＶＲＮＡ。４９例感染肝组织分为两组：丁肝组２３例；单纯乙肝组２６例，ＨＢＶＤＮＡ的检出率丁肝组（７８．２６％）与乙肝组（７６．９２％）无统计学差异；而ＨＤＶＮＡ的检出率丁肝组（６０．８７％）明显高于乙肝组（１５．３８％）。ＨＢＶＤＮＡ可见于受染肝细胞的胞核或胞浆内，而ＨＤＶＲＮＡ绝大部分见于肝细胞胞核。两种病毒核酸阳性细胞在肝组织中的分布特点大致相同：弥漫或散在地分布于肝小叶或假小叶内，或局灶性分布于小叶周边。ＨＤＶＲＮＡ阳性的肝组织都或多或少地同时存在ＨＢＶＤＮＡ。同一例肝组织中，ＨＢＶＤＮＡ阳性细胞从数量和颗粒密度上似略高于ＨＤＶＲＮＡ。将乙肝组和丁肝组两组病人肝内ＨＢ－ｓＡｇ、ＨＢｃＡｇ和ＨＢＶＤＮＡ及血清ＨＢｅＡｇ作了比较，各指标阳性率虽有差异，但均无统计学意义。因此，未发现ＨＤＶ感染对ＨＢＶ的复制有明显抑制作用。此结果对以往用血清学或免疫组化方法对ＨＤＶ的研究有所补充和深入，亦可为研究其它类型病毒性肝炎之间的重叠感染所借鉴。     

Investigating the prevalence and clinical effects of hepatitis delta viral infection in Taiwan

PurposeTo clarify and investigate the prevalence and clinical impact of hepatitis D virus (HDV) infection in Taiwan's communities.MethodsHDV infection in patients with chronic hepatitis B viral (HBV) infection was examined using an anti-HDV antibody in Yonghe Cardinal Tien Hospital (YCTH), a district hospital in Taiwan. Clinical characteristics of anti-HDV-positive and anti-HDV-negative patients were collected and compared. These characteristics were also compared with the data collected from a medical center. Continuous variables and confounding factor adjustments were compared using the analysis of covariance method, whereas categorical variables were compared using the logistic regression method.ResultsA total of 346 patients with chronic HBV infection were assessed from 2018 to 2019. Among them, 4 (1.15%) were positive for anti-HDV. The clinical, virological, and biochemical characteristics were similar between anti-HDV-positive and anti-HDV-negative groups. None of the four patients was positive for serum HDV RNA. Another 18 anti-HDV-positive patients were identified from National Taiwan University Hospital (NTUH). The clinical, virological, and biochemical characteristics of anti-HDV-positive patients from YCTH and NTUH were also similar.ConclusionThe prevalence of HDV and the serum HDV RNA-positive rate were low in district hospitals in Taiwan. Coexisting HDV infection did not influence the clinical manifestation of patients with chronic HBV infection in Taiwan. However, because the number of HDV RNA cases was very small, our findings may not be conclusive. Besides, since the sensitivity of current anti-HDV kit is not 100%, more sensitive methods are needed to achieve reliable prevalence data.