应用基因工程菌表达的丁型肝炎抗原(HDAg)制备抗-HD酶联免疫试剂及其应用

采用基因工程技术使国外引进的HDAg基因克隆株在大肠杆菌中转化和表达。以这种工程菌提取的HDAg为抗原,建立了检测血清中抗-HD的ELISA试剂,采用竟争法测定。与爱尔兰Noc-tech及美国Abbott两公司的同类试剂盒相比,前者57份血清检测结果,2份阳性和55份阴性全部符合;后者55份血清中2份阳性和53份阴性,亦均完全符合。对本院HBsAg阳性各类型肝炎或携带者血清测定抗-HD的检出率为6.38％(25/392);对国内8省市同类病例检出率为0.69％(5/729)。临床及流行病学调查表明本试剂特异性、敏感性均良好。对菌产HDAg的特性和酶联试剂的特点进行了分析和讨论。     

392例HBsAg阳性急慢性肝炎和携带者血清δ系统的研究

本文应用国产丁型肝炎病毒(HDV)酶联免疫试剂对392份HBsAg阳性肝炎和携带者血清进行了抗-HD、抗-HDIgM和HDAg测定,HDV总感染率为10.7％(42/392)。其中以重型肝炎组HDV标志检出率最高,达27.8％,然后依次为慢性肝病(15.5％)和急性乙肝(5.3％),HBsAg阳性携带者无1例阳性。本研究资料提示HDV感染对HBV感染在加重肝损害、促进肝脏炎症进展及慢性化方面均起十分重要作用。HDV标志与HDV RNA的测定亦有较高的符合率(96.9％)。     

平湖市丁型肝炎病毒感染状况的调查

目前,在我国乙型肝炎患者中丁型肝炎的检出率很高,最高可达30%,为了解丁型肝炎在乎湖市感染情况,我们对76名乙肝携带者进行了抗-HD和HDAg检测,结果如下。材料和方法1．检测对象,在健康体检中乙肝表面抗原阳性者分离血清1ml,放置-20℃冻存待检。2．检测方法:ALT和乙肝两对半试剂均由上海科华所提供,抗-HD和HDAg试剂为北京302医院提供,操作按说明书进行。结果1．抗-HD和HDAg检测结果,76名表抗阳性者中抗-HD阳性3人,阳性率为3.94%,HDAg阳性5人,阳性率为6.58%,无双阳性,总HDV感染率为10.53%。     

抗丁型肝炎病毒lgM抗体检测试剂盒的制备及应用

本文报告用基因工程生产的HDAg和高效价的抗-HD阳性血清,成功地制备了检测抗-HDIgM的ELISA试剂盒。同国外同类产品比较,其灵敏度为93.9％,特异性为100％。本试剂与甲乙肝抗原无交叉反应,不受类风湿因子的干扰。经临床应用及国内8省市的流行病学调查,均取得了较满意结果。     

HDV重组抗原的测定及与其它抗原的比较

[目的]为了研究重组HDV抗原的性质及与其它HDV抗原的差别。[方法]本文用ELISA方法对重组HDAg进行了测定,并与肝脏提取的HDAg及美国HDAg进行了比较实验。[结果]重组HDAg能与抗-HD特异性结合,中和抑制试验平均抑制率为86％,该抗原在37℃放置1～7d稳定性良好。[结论]三种抗原对抗-HD阳性血清和HBsAg阳性血清的检测结果一致,表明基因重组 HDAg可以替代诊断试剂中的其他HDV抗原。     

急性δ感染时HDV RNA:血清学特征和其他HDV感染标记的相互关系

急性HDV感染的血清学诊断是基于HDAg和/或抗-HD的测定。在HDV和HBV联合感染时,HDV标记往往呈一过性,而慢性HBsAg携带者重叠HDV时,多呈慢性感染并伴血清总抗-HD和IgM抗-HD阳性以及肝内HDAg。最近,应用分子杂交技术测定HDV感染患者的血清HDV RNA,旨在明确其临床意义以及观察与急性或慢性HDV感染的其他标记(如肝内HDAg和血清IgM抗-HD)的相互关系。     

住院乙型肝炎病人血清丁型肝炎病毒感染标志物检测

为了解乙型肝炎病人丁型肝炎病毒感染情况,对在我院住院的乙肝病人,进行HDAg和抗-HD检测.1对象与方法被检者为1998年4～10月在我院住院的乙肝病人,血清HBsAg均阳性.血清HDAg和抗-HD检测用ELISA法.试剂为北京科卫临床诊断试剂厂产品.     

IgM抗-HD为慢性δ感染的标志

慢性δ肝炎的诊断依据通常是血清中存在高滴度抗δ总抗体(抗-HD)和/或肝细胞中存在δ抗原(HDAg)。IgM 抗-HD 可在部分急性δ病毒(HDV)感染期病人中短暂出现,并已成为慢性感染的一项敏感标志。作者测定了一组抗-HD 阳性慢性肝病病人的抗体和肝内 HDAg,并研究了这两项标志与肝损害的程度和严重性的关系。共观察36例 HBsAg 阳性及组织学证实为慢性肝病和高滴度抗-HD 抗体(>1∶     

慢性丁型肝炎:免疫印迹法检测血清中HDAg及其与其他HDV复制标志的关系

慢性丁型肝炎病毒(HDV)感染的诊断通常是根据肝组织内发现丁型肝炎病毒抗原(HDAg)及血清中有高滴度的HDV抗体(抗-HD),极少情况下可用ELISA检测到血清HDAg。1987年以来,有少数学者开始应用免疫印迹法检测慢性HDV感染病人血清HDAg,并用此法证实有两条抗原带,分子大小分别为24kD和27kD,近5年又证实了急、慢性HDV感染病人血清中HDV RNA的存在并且与肝组织内     

HBV携带者中HDV抗原抗体的检测

作者于1991年1～8月对追踪观察的 328例HBV携带者(其中266例是无症状带毒者,48例是慢性肝炎,14例是肝硬变和肝癌)用ELISA法分别测定其血中抗-HD、IgM抗-HD及HDAg,并用RIA法对抗-HD阳性者作了进一步证实。肝组织中的HDAg用酶抗体法检测。结果:在328例HBV携带者中检出6例(1.8％)抗-HD阳性者,其中62例慢性肝病者中检出5例(8.1％),266例无症状带毒者中检出1例(0.4％),两组间的检出     

丁型肝炎系列酶免诊断试剂的研制及应用

为解决丁型肝炎诊断试剂短缺的问题，本研究采用实验感染ＨＤＶ的土拨鼠肝脏和丁型肝炎病人血清，提取纯化丁型肝炎病毒抗原和抗体，制备了检测ＨＤＡｇ、抗－ＨＤ、ＩｇＭ抗－ＨＤ的系列酶免诊断试剂。该试剂与进口同类试剂相比较，符合率达１００％。其特异性好，灵敏度较高。对不同类型乙型肝炎病人血清检测，ＨＤＡｇ、抗－ＨＤ及ＩｇＭ抗－ＨＤ阳性率和总阳性率分别为２．９９％、３．３６％、３．７３％和７．２８％。该试剂的研制为丁型肝炎病毒感染的临床诊断及流行病学研究提供了可靠的实验手段。     

Unexpected low prevalence of delta antibodies in the east Amazon region and S?o Paulo: evidence for regional differences in the epidemiology of delta hepatitis virus within Brazil

Antibodies (anti-HD) to hepatitis delta virus (HDV) were tested by radioimmunoassay in 207 human serum samples from the eastern Amazon (states of Pará and Amapá) and S?o Paulo, Brazil. 42 Amazon HBsAg asymptomatic carriers were negative for anti-HD. 84 S?o Paulo HBsAg asymptomatic carriers were also negative. Among the 81 HBsAg patients from S?o Paulo with different liver diseases, only one had anti-HD. Liver biopsy of this chronic active hepatitis case was positive for HBsAg, HBcAg and HDAg in liver, by an immunoperoxidase technique. The low prevalence of HDV infections in S?o Paulo and eastern Amazon was unexpected and contrasts with the recent reports of high prevalence in the western Amazon region. Such regional differences emphasize the need for extensive and precise worldwide epidemiological studies of HDV.     

丁型肝炎病毒感染的血清流行病学观察

1987年4月至1988年10月间,本文应用酶联吸附法(EIA)对石家庄地区Ml例乙型肝炎病毒感染者进行了抗-HD的检测,共发现35例阳性,阳性率12.92%,其中男性阳性率14.06%(27/102),女性为10.13%(8/79),男女间差异无统计学意义(P>0.05),提示石家庄地区可能为丁型肝炎病毒感染的高发区.在这些人群中,慢性活动性肝炎、慢性迁延性肝炎和肝硬化的抗-HD阳性率明显高于HBsAg携带者,但三者相互间差异无统计学意义,表明合并或重香感染HDV对乙肝慢性化及肝硬化的形成具有重要的意义。本研究证明在乙型肝炎病毒感染人群中丁型肝炎病毐与年龄、性别、职业等因素关系不密切。     

Immunization of woodchucks (Marmota monax) with hepatitis delta virus DNA vaccine

We investigated the DNA immunization approach in order to induce a protective immune response against hepatitis delta virus (HDV) superinfection of chronically woodchuck hepatitis virus (WHV) infected woodchucks. The animals were immunized with an expression vector encoding HDAg by gene gun. T cell and humoral immune responses induced by this protocol were determined and compared with those induced by HDAg immunization using a CpG oligonucleotide as an adjuvant. After immunization the woodchucks were challenged with 10⁶ genome equivalents of HDV. The protein immunization with HDAg induced good humoral and T helper cell responses in the woodchucks, but did not protect them from HDV superinfection. The DNA immunized woodchucks were also not protected from HDV superinfection, however, the course of infection was modified: HDV viremia occurred later, the typical fluctuation of the HDV RNA titer with several peaks was absent, and antibodies to HDV were not detectable.     

河南省人群丁型肝炎病毒感染的血清流行病学调查研究

为了了解河南省丁型肝炎病毒的人群感染情况，从１９９１－１９９３年对河南省十三个地区１１８２例ＨＢｓＡｇ阳性的各型乙肝病人及无症状ＨＢｓＡｇ携带者血清标本进行ＨＤＡｇ、抗ＨＤＶ和ＩｇＭ－抗－ＨＤＶ三项标志检测。结果表明，河南省人群ＨＤＡｇ、抗－ＨＤＶ、ＩｇＭ－抗－ＨＤＶ的总感染率分别为３．０％、３．５％、３．５％、８．１％。     

Immunization of woodchucks (Marmota monax) with hepatitis delta virus DNA vaccine

We investigated the DNA immunization approach in order to induce a protective immune response against hepatitis delta virus (HDV) superinfection of chronically woodchuck hepatitis virus (WHV) infected woodchucks. The animals were immunized with an expression vector encoding HDAg by gene gun. T cell and humoral immune responses induced by this protocol were determined and compared with those induced by HDAg immunization using a CpG oligonucleotide as an adjuvant. After immunization the woodchucks were challenged with 10⁶ genome equivalents of HDV. The protein immunization with HDAg induced good humoral and T helper cell responses in the woodchucks, but did not protect them from HDV superinfection. The DNA immunized woodchucks were also not protected from HDV superinfection, however, the course of infection was modified: HDV viremia occurred later, the typical fluctuation of the HDV RNA titer with several peaks was absent, and antibodies to HDV were not detectable.     

重庆部分地区HDV感染的调查

本文收集了重庆部分地区乙肝患者ＨＢｓＡｇ无症状携带者血清标本２４２份，用ＥＬＩＳＡ法检测抗－ＨＤ，２４份阳性，阳性率为９．９１％，与其它地区比较重庆部分地区ＨＤＶ感染率较高，且郊区的阳性率明显高于市区，此结果符合以前曾报告的ＨＤＶ在我国呈地方性分布的论述。原发性肝癌病人，急、慢乙型肝炎病人和ＨＢｓＡｇ无症状携带者中，以原发性肝癌病人抗－ＨＤ阳性率较高，提示ＨＤＶ感染可能与原发性肝癌的致病有关。     

The prevalence of delta virus infection in chronic liver disease in Egyptian children in comparison with some other countries

BACKGROUND: Hepatitis Delta Virus (HDV) infection has a worldwide distribution. The aim of this study was to establish its prevalence in Egyptian children with chronic liver disease, and its role in the development and progress of hepatic illness. METHODS: A prospective study of 45 Egyptian children who had liver cirrhosis (n = 24) or chronic hepatitis (n = 21) was done. They were consecutively chosen. Their ages ranged from 2-15 years (median = 5). Serological studies were performed to detect the presence of hepatitis B surface antigen (HBs Ag) and HDV infection IgG antibody (IgG anti-HD). RESULTS: Anti-delta antibody (IgG anti-HD) was detected in only four children with hepatic cirrhosis and non of the chronic hepatitis, with an overall prevalence of 8.9% (4/45). Three of them (75%) were hepatitis B surface antigen (HBs Ag) negative. Significant statistical association between delta infection and the state of hepatic illness was detected (p < 0.05). Whereas HBs Ag was detected in 54% (13/24) of liver cirrhosis and 52% (11/21) of chronic hepatitis, with an overall prevalence of 53% (24/45). There was no statistically significant association between HBs Ag positivity and state of hepatic illness. CONCLUSIONS: The prevalence of HDV infection is 8.9% of Egyptian children with chronic liver disease. HDV infection in children is associated with advanced chronic liver disease.     

Delta infection in eastern Sicily

Sera from 619 HBsAg+ subjects living in eastern Sicily, consecutively collected from 1975–1985, were tested for markers of delta virus (HDV) infection: delta antigen (HDAg), antibodies to delta (anti-HDIg), and also for antibodies to HBcore of IgM type (anti-HBcIgM) and for the system HBe-anti-HBe. The subjects included 210 asymptomatic carriers, 238 patients with acute hepatitis and 171 patients with chronic liver disease.HDAg was not found in any of the samples. Anti-HD was found in 28/171 (16.3%) patients with chronic liver disease, in 13/210 (6%) asymptomatic HBsAg carriers and in 13/238 (5.4%) patients with acute hepatitis. None of our patients were drug addicts. One had a history of blood transfusion, and nine came from the same family unit.The prevalence of HDV infection in eastern Sicily is lower than in other areas of Sicily possibly because of the lower percentage of HBsAg carriers in the local population. Parenteral transmission of HDV does not seem to play a major role in our area, while the familial clustering suggests close body contact as an important way of spread.