大理地区猪源甲型、乙型、戊型肝炎病毒血清流行病学调查

目的:调查大理地区猪体内甲、乙、戊型肝炎病毒的感染情况。方法:采用酶联免疫吸附实验(ELISA)检测屠宰生猪血清标本甲肝血清标志物(HAV-lgG和HAV-lgM)、乙肝血清标记物(HBsAg,HBsAb,HBeAg,HBeAbHBcAb)、戊肝血清标志物(HEV-lgG和HEV-lgM)。结果:HAV、HBV和HEV的感染率分别为:91%(162/178),93.9%(185/197),23.6%(42/178);HAV和HBV二重感染率为67.97%(121/178);HAV、HBV和HEV三重感染率为20.2%(36/178);HAV和HEV二重感染率为1.7%(3/178);HBV和HEV二重感染率为2.2%(4/178)。结论:甲、乙、戊型肝炎病毒在猪体内存在感染,同时发现在猪体内甲、乙型肝炎病毒重叠感染常见。     

Prevalence of hepatitis A virus,hepatitis B virus,hepatitis C virus,hepatitis D virus and hepatitis E virus as causes of acute viral hepatitis in North India: A hospital based study

Context: Acute viral hepatitis (AVH) is a major public health problem and is an important cause of morbidity and mortality. Aim: The aim of the present study is to determine the prevalence of hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV) and hepatitis E virus (HEV) as causes of AVH in a tertiary care hospital of North India. Settings and Design: Blood samples and clinical information was collected from cases of AVH referred to the Grade I viral diagnostic laboratory over a 1-year period. Subjects and Methods: Samples were tested for hepatitis B surface antigen, anti-HCV total antibodies, anti-HAV immunoglobulin M (IgM) and anti-HEV IgM by the enzyme-linked immunosorbent assay. PCR for nucleic acid detection of HBV and HCV was also carried out. Those positive for HBV infection were tested for anti-HDV antibodies. Statistical Analysis Used: Fisher’s exact test was used and a P < 0.05 was considered to be statistically significant. Results: Of the 267 viral hepatitis cases, 62 (23.22%) patients presented as acute hepatic failure. HAV (26.96%) was identified as the most common cause of acute hepatitis followed by HEV (17.97%), HBV (16.10%) and HCV (11.98%). Co-infections with more than one virus were present in 34 cases; HAV-HEV co-infection being the most common. HEV was the most important cause of acute hepatic failure followed by co-infection with HAV and HEV. An indication towards epidemiological shift of HAV infection from children to adults with a rise in HAV prevalence was seen. Conclusions: To the best of our knowledge, this is the first report indicating epidemiological shift of HAV in Uttar Pradesh.     

TT病毒与肝炎关系的临床流行病学研究

目的 对闽南地区各种肝炎患者、健康体检者、义力献血员和肝癌患者共４８０例从临床流行病学角度探讨ＴＴ病毒（ＴＴＶ）的致生及其与各种肝炎的关系。方法 采用巢式ＰＣＲＩｇＧ,用ＥＰＩ ＩＮＦＯ６．０软件进行统计分析。结果 ４８０名研究对象中ＴＴＶＤＮＡ的总检出率为２３．９６％。各种胆炎患者的ＴＴＶ总阳性率为２３．９４％,肝癌患者的ＴＴＶ阳性率为２０．６９％,而健康者的ＴＴＶ阳性率为２４．８４％,义务献血     

用原位酶联免疫吸附法检测甲型肝炎病毒

目的　采用一种更加简单和敏感的检测方法对甲型肝炎病毒(HAV)进行检测。方法　用甲型肝炎病毒疫苗株H2M20K株和野毒株合34感染KMB17二倍体细胞,于96孔微量细胞板上培养,并在原位用HAV特异的单克隆抗体与单层细胞上的病毒结合,辣根过氧化物酶标记的抗鼠IgG作为指示,以细胞对照A值(OD)均数的18倍判为阳性。结果　病毒增殖动态显示,高峰期为20～25d,比ELISA终点滴定法提前4～6d;两种方法同时做13个样品的感染性滴度比较,结果差异无显著意义(t=1.13,P>0.05)。原位EIA较好地指示中和试验的结果;直接从12份粪便中分离病毒,经第一代培养检测有5份阳性。结论　EIA敏感性与ELISA法相似,由于原位EIA直接检测病毒细胞系统,有操作简单,重复性好等优点,可取代ELISA终点滴定法,同时也可望用于对其他病毒细胞系统的检测。     

Survival of hepatitis A and E viruses in soil samples

Survival of hepatitis A virus (HAV) and hepatitis E virus (HEV) in soil samples spiked with respective viruses was analysed using real-time PCR. Virus-spiked soil samples were incubated at environmental temperature (ET) and 37°C and processed weekly. Both HAV and HEV were less stable at fluctuating ET than at 37°C. Of the 403 soil samples collected in the vicinity of Mutha river, India, 19.1% and 4.9% were found to be contaminated with HAV and HEV, respectively.     

检测病毒性肝炎患者血清中SEN病毒及其临床意义

目的：检测病毒性肝炎患者血清中SEN病毒D和H(SENV-D、SENV-H),并探讨其临床意义。方法：采用巢式聚合酶链反应法（nPCR）检测甲、乙、丙、戊型肝炎和非甲－戊型肝炎患者血清中SENV-D和SENV-H DNA。结果：在180例病毒性肝炎患者血清中，SENV-D和SENV-H检出率分别为17.2%(31/180)和5.6%(10/180)，总检出率为18.3%(33/180)、甲、乙、丙、戊型肝炎患者的SENV-D/H检出率高于非甲－戊型肝炎患者。从甲、乙、丙、戊型肝炎和非甲－戊型肝炎患者分离的SENV-D/H核苷酸序列，与SENV-D/H原型株比较，其同源性在94％以上。甲、乙、丙和戊型肝炎患者有无SENV-D/H合并感染，其血清生化学指标无明显差异。结论：SENV-D/H可能不是非甲-戊型肝炎的病原，甲、乙、丙和戊型肝炎患者合并感染SENV-D/H并不加重病情。     

庚型肝炎病毒IgM抗体检测方法的建立及其临床意义

目的建立一种早期、快速诊断庚型肝炎的血清学检测方法。方法以辣根过氧化物酶标记庚型肝炎病毒（ＨＧＶ）多肽ＮＳ３,ＮＳ５区段抗原,建立了捕获酶联免疫吸附试验（ＥＬＩＳＡ）,用于检测血清中ＨＧＶＩｇＭ抗体。结果本法不受特异性ＩｇＧ的竞争和类风湿因子的干扰;与其它致肝炎的病毒（ＨＡＶ、ＨＢＶ、ＨＣＶ、ＨＥＶ、ＣＭＶ、ＥＢＶ）无交叉反应。检测４６例非甲、乙、丙、戊型肝炎患者血清,抗－ＨＧＶＩｇＭ阳性１４例,阳性率３０．４３％,其中,６例同时为ＨＧＶＲＮＡ阳性,阳性符合率为４２．８６％（６／１４）,检测１２例庚型肝炎病人双份血清,其中,急性期血ＨＧＶＩｇＭ抗体均为阳性。结论该法检测ＨＧＶＩｇＭ抗体特异性强,敏感性高,且简便快速,适用于临床对庚型肝炎新近感染的早期诊断,有推广应用价值。     

Autochthonous hepatitis E in Southwest England: a comparison with hepatitis A

The incidence of hepatitis A is falling. In contrast, autochthonous hepatitis E is an emerging infection in developed countries. The objective of this study was to compare both laboratory-confirmed cases of hepatitis A and autochthonous hepatitis E over a 2-year period in Cornwall and Devon and anti-hepatitis A virus (HAV) IgG and anti-hepatitis E virus (HEV) IgG seroprevalence in blood donors. The databases of microbiology laboratories in Cornwall and Devon were searched for the number of diagnostic HEV and HAV assays performed during 2005-2006 and the number of confirmed cases of acute hepatitis A and hepatitis E detected. Patients were followed up until recovery or death. Sera from 500 blood donors from the regional centre were tested for HEV and HAV IgG. In total, 28 cases of autochthonous hepatitis E were identified from 838 assays, and 20 cases of hepatitis A were identified from 4503 assays. Compared to hepatitis A cases, patients with hepatitis E were older (mean age 61 vs. 45 years, P = 0.003), less likely to present in winter (P = 0.028) and had more complications (five vs. one). The IgG seroprevalence rates in blood donors were 45% for HAV and 16% for HEV. There was no relationship between HAV and HEV IgG seropositivity. Autochthonous hepatitis E may be more common than hepatitis A, affects older patients, is less likely to occur in winter and may be associated with more complications. Patients with acute hepatitis, whatever their age or travel history, should be tested for HEV.     

Coinfection of hepatitis A virus genotype IA and IIIA complicated with autoimmune hemolytic anemia,prolonged cholestasis,and false-positive immunoglobulin M anti-hepatitis E virus: a case report

A 37-year-old male presented with fever and jaundice was diagnosed as hepatitis A complicated with progressive cholestasis and severe autoimmune hemolytic anemia. He was treated with high-dose prednisolone (1.5 mg/kg), and eventually recovered. His initial serum contained genotype IA hepatitis A virus (HAV), which was subsequently replaced by genotype IIIA HAV. Moreover, at the time of development of hemolytic anemia, he became positive for immunoglobulin M (IgM) anti-hepatitis E virus (HEV). We detected HAV antigens in the liver biopsy specimen, while we detected neither HEV antigen in the liver nor HEV RNA in his serum. This is the first report of hepatitis A coinfected with two different genotypes manifesting with autoimmune hemolytic anemia, prolonged cholestasis, and false-positive IgM anti-HEV.     

Differences in capabilities of different enzyme immunoassays to detect anti-hepatitis E virus immunoglobulin G in pigs infected experimentally with hepatitis E virus genotype 3 or 4 and in pigs with unknown exposure

Hepatitis E virus (HEV), a major cause of acute viral hepatitis in humans in many developing countries, is highly prevalent in the pig population worldwide. The objective of this study was to assess the capability of three porcine prototypes of a human enzyme-linked immunosorbent assay (ELISA), an in-house ELISA and a line-immunoassay (LIA) to detect anti-HEV antibodies in pigs infected experimentally with HEV (n = 57), known to be negative for HEV infection (n = 27), or with unknown exposure to HEV infection (field samples, n = 90). All 27 samples from non-infected pigs were negative with all five assays. The earliest detection of anti-HEV antibodies occurred at 14 days post-inoculation (dpi) with four of five assays. From 42 dpi, all samples from infected pigs were detected correctly as anti-HEV positive. Kappa analysis demonstrated substantial agreement among tests (0.62-1.00) at 14 dpi and complete agreement (1.00) at 56 dpi. The overall area under the curve for all quantitative tests as determined by receiver operator characteristic analysis ranged from 0.794 to 0.831 indicating moderate accuracy. The results showed that all five assays can detect anti-HEV IgG antibodies accurately in pigs infected experimentally with HEV. In field samples, a higher prevalence of anti-HEV IgG was found in breeding herds than in growing pigs (100% versus 66.7-93.9%). These serological assays should be very useful in veterinary diagnostic labs for HEV diagnosis in swine.     

1995至2000年北京地区散发性急性肝炎患者肝炎病毒抗体的检测与分析

目的 了解北京市地区散发性肝炎患者甲型、乙型、丙型和戊型肝炎病毒感染型别分布及重叠感染。方法 用EIA法检测1995斫2月至2000年12月北京地区散发性急性肝炎患者抗－HAVIgM、HBsAg／抗－HBcIgM、抗－HCVIgM/IgG和抗－HEVIgM/IgG。结果 214例散发性急性肝炎患者血清,抗甲、乙、内和戊型肝炎病毒IgM总阳性数155例,戊型肝炎76例。有9名患者检出2种肝炎病毒抗原或抗体阳性,其中在3名肝硬化患者和2名静脉吸毒者同时检测到HBV和HCV抗原或抗体,1例HBsAg阳性者检测到抗－HAVIgM,3例－HCVIgG阳性者中分别检测到2例抗－HBVIgM和1例抗－HEVIgM。肝炎病毒重叠感染的9名患者年龄在31岁至49岁之间。结论 北京地区散发性急性肝炎78％是由消化道传染的甲戊型肝炎病毒引起,戊型肝炎在四种肝炎中位居首位,其次为甲型肝炎、乙型肝炎和丙型肝炎。肝炎病毒重叠感染多见乙、丙型肝炎病毒合并感染或慢性乙、丙型肝炎患者合并甲型或戊型肝炎病毒感染。在我国对散发性病毒性肝炎的预防应引起高度重视。     

Unrelatedness of factor VIII-derived non-A/non-B hepatitis and hepatitis B virus

A DNA hybridization assay was used to detect hepatitis B virus (HBV)-specific DNA sequences in extracted sera obtained from chimpanzees infected with HBV, hepatitis A virus (HAV), and a factor VIII-derived non-A/non-B (NANB) agent. The results did not reveal any HBV-DNA homology with sera obtained from animals infected with HAV or factor VIII-derived NANB. Sera obtained from two HBV-infected chimpanzees demonstrated that HBV-specific DNA could be detected during the acute phase of the disease. In addition, an HBV-specific DNA-dependent DNA polymerase assay did not demonstrate any statistically significant activity in 12 of 12 NANB acute-phase specimens or in 6 of 6 NANB chronic-phase specimens. These results suggest that the factor VIII-derived NANB agent is unrelated to HBV.     

Distinct changing profiles of hepatitis A and E virus infection among patients with acute hepatitis, patients on maintenance hemodialysis and healthy individuals in Japan

To compare the epidemiologic profiles of hepatitis A virus (HAV) and hepatitis E virus (HEV) infections in Japan, the prevalence of clinical or subclinical HAV and HEV infections was investigated serologically and molecularly among 128 consecutive patients (age, mean +/- standard deviation, 37.5 +/- 14.7 years) who contracted acute hepatitis between 1989 and 2005 in a city hospital, and among 416 hemodialysis patients (60.1 +/- 12.6 years) and 266 medical staff members (34.6 +/- 11.4 years) at the same hospital, using stored periodic serum samples collected since the start of hemodialysis or employment, respectively. Between 1989 and 1995, among 93 patients with acute hepatitis, 51 (54.8%) were diagnosed with hepatitis A and only one patient with hepatitis E. Between 1996 and 2005, however, among 35 patients, only 3 (8.6%) were diagnosed with hepatitis A and 2 (5.7%) with hepatitis E. Although subclinical HEV infection was recognized in four hemodialysis patients (one each in 1979, 1980, 1988, and 2003) and two medical staff members (1978 and 2003) in previous studies, none of the 191 hemodialysis patients who had been negative for anti-HAV at the start of hemodialysis contracted HAV infection during the observation period of 7.6 +/- 6.4 years. Only one (0.4%) of the 246 medical staff members who had been negative for anti-HAV at the start of employment acquired hepatitis A during the observation period of 7.9 +/- 8.0 years: none had subclinical HAV infection. Clinical or subclinical HEV infection has occurred rarely during the last three decades, while HAV infection has markedly decreased at least since 1996.     

Association of hepatitis E virus with an outbreak of hepatitis in Pakistan: serologic responses and pattern of virus excretion.

Hepatitis E virus (HEV), a positive-strand RNA agent, has been associated with enterically transmitted non-A, non-B hepatitis in Asia, Africa, and Mexico. To evaluate the role of HEV in an outbreak of hepatitis in Pakistan, we used immune electron microscopy to detect 1) antibody to HEV, for evidence of infection, and 2) virus, to determine the pattern of HEV excretion. Paired sera from 2 patients were assayed for antibody by using reference HEV: one seroconverted, an atypical finding for HEV infections; the other had high levels of anti-HEV in both sera. Virus particles with the size (29 x 31 nm) and morphology of HEV were detected in feces from 10 of 85 patients and serologically identified as HEV by using reference antibodies from an HEV-infected chimpanzee. One of these HEV-containing specimens was collected 9 days before the onset of jaundice; it was among feces from 38 outpatients with nonspecific symptoms and biochemical hepatitis, 12 of whom subsequently developed jaundice. The other 9 feces with HEV were among 36 collected within 7 days of the onset of acute icteric hepatitis; all 11 feces from days 8 to 15 were negative for HEV. Fecal concentrations of HEV appeared to be lower than those of many enteric viruses: only one specimen contained as many as 5 particles per EM grid square. It is concluded that HEV was etiologically associated with the epidemic and was predominantly excreted at very low levels during the first week of jaundice.     

抗甲肝病毒人源基因工程全抗体分子在杆状病毒中的表达

目的 探讨人源抗甲型肝炎病毒全抗体分子在杆状病毒中的表达。方法 将获得的人源抗甲肝病毒中和性抗体Fab段基因克隆入含信号肽及Fc的杆状病毒表达载体中并在杆状病毒细胞中表达。结果 获得了中和性人源抗甲肝病毒全抗体分子的表达产物并进行了纯化，轻重链表达产物位置大小正确，HAFc16抗体能与具有中和活性的鼠抗甲肝病毒单克隆抗体产生竞争抑制反应，并能在体外中和甲肝病毒，另一株HAFc78抗体同样具有体外中和甲肝病毒的活性，但系抗不同位点的抗体。结论 获得的人源抗甲肝病毒全抗体分子表达产物具有很好的体外中和甲肝病毒的活性，且为抗不同位点的抗体，为这些抗体的进一步开发及应用打下了基础，为防止甲型肝炎暴发流行提供应急措施。     

Genotypic characterization of symptomatic hepatitis E virus (HEV) infections in Egypt

BackgroundHepatitis E virus (HEV) is a common cause of acute viral hepatitis (AVH) in many developing countries. In Egypt, HEV seroprevalence is among the highest in the world; however, only a very limited number of Egyptian HEV sequences are currently available.ObjectivesThe objectives were to determine the HEV genotype(s) currently circulating in Egypt.Study designAVH patients without serologic evidence of hepatitis A, B, and C viruses were evaluated for possible HEV infection using serologic assays for anti-HEV IgM and anti-HEV IgG and real-time PCR for HEV RNA. Stool suspensions from suspected cases were inoculated into rhesus macaques to confirm the presence of HEV. Sequence analysis was utilized to determine HEV genotype.ResultsOf 287 subjects with AVH enrolled, 58 had serologic evidence of acute HEV infection. Stool samples for two of these patients were repeatedly positive for HEV RNA by real-time PCR. Macaques experimentally inoculated with these human stools also developed viremia. Sequence analysis of open reading frame (ORF) 1 demonstrated that these isolates belonged to HEV genotype 1 and were 3.9–9.5% divergent from other genotype 1 isolates. ORF2 was 5.3–8.7% divergent from previously reported Egyptian isolates.ConclusionsThis study strongly suggests that genotype 1 HEV related to other North African isolates is circulating in acute symptomatic patients in Egypt. Further evaluation of genotypic variability is underway in this highly endemic cohort and is considered an important component of our increased understanding of HEV pathogenesis.     

甲肝病毒特异性TaqMan荧光定量PCR法检测甲肝病毒核酸的研究

目的 建立甲肝病毒(HAY)特异性TaqMan荧光定量PCR检测方法,并对血清等样品中的HAV进行检测.方法 根据参考文献,选取HAV基因组保守区5'-NCR区引物和探针,建立特异性强、敏感度高、稳定性好的TaqMan HAV Real-time RT-PCR检测体系并应用于甲肝患者血清等病毒核酸的检测.结果 本研究建立的方法能够特异检测出样品中的甲肝病毒,其灵敏度介于每反应0.1CCID50至0.01CCID50之间.同一样本重复检测3次,批内样本Ct值的变异系数最大2.0％,批间样本Ct值的变异系数最大2.6％.急性期甲型肝炎患者血清中甲肝病毒为5.18×102～4.93×107拷贝/ml.结论 本实验建立的HAV Real-time RT-PCR检测方法具有特异、灵敏、快速等优点,可成功应用于临床样本等甲肝病毒的病原学检测.     

Comparison of hepatitis A and E virus infections among healthy children in Mongolia: evidence for infection with a subgenotype IA HAV in children

To compare the epidemiologic profiles of hepatitis A virus (HAV) and hepatitis E virus (HEV) infections in children in Mongolia, the prevalence of HAV and HEV infections was investigated serologically and molecularly among 717 apparently healthy individuals of 0-20 years of age (mean +/- standard deviation, 8.6 +/- 4.9 years) using serum samples obtained between October 2005 and January 2006. Total antibody against HAV (anti-HAV [total]) was detected in 494 (68.9%) of the 717 subjects, while IgG antibody against HEV (anti-HEV IgG) was detected in only five subjects (0.7%) (P < 0.0001). All five subjects who had anti-HEV IgG, were negative for anti-HEV IgM and HEV RNA. Anti-HAV was detectable in 24 (75.0%) of the 32 infants aged 7 days to 6 months, but not in any of the 8 infants aged 7 to <12 months. The prevalence of anti-HAV was 19.5% (17/87) in the age group of 1-3 years, and it increased to 50.0% (69/138) in the age group of 4-6 years, and further to 81.4% (105/129) in the age group of 7-9 years. Of note, 97.2% of the subjects in the age group of 16-20 years had anti-HAV. The presence of HAV RNA was tested in all 717 subjects, and three children of 1, 4, or 8 years of age were found to have detectable HAV RNA (subgenotype IA). No subject had a history of hepatitis or jaundice. In conclusion, HEV infection was uncommon, but HAV infection lacking overt clinical features was prevalent among children in Mongolia.     

Prevalence of antibodies to hepatitis E virus in residents of a district in Havana, Cuba

A seroepidemiological study of hepatitis E virus (HEV) infection was conducted in a district of Havana, where hepatitis A virus (HAV) is considered endemic. The levels of anti-HEV antibodies were evaluated by enzyme-linked immunosorbent assay (ELISA) based on the recombinant protein GST-ORF2.1. Anti-HEV antibodies were detected in 11 of 209 (5.3%) of serum samples, compared to 71.3% for anti-HAV antibodies. No risk factors reported previously for HEV infection showed a significant association with the presence of anti-HEV antibodies, whereas anti-HAV antibodies were strongly associated with increasing age. HEV may be considered endemic in this area and is likely to have a significant clinical impact.     

Development of a fluorescent microbead-based immunoassay for the detection of hepatitis E virus IgG antibodies in pigs and comparison to an enzyme-linked immunoassay

Swine hepatitis E virus (HEV) is a zoonotic virus and pigs are considered as an important reservoir. Swine HEV infection is widespread and most pig herds are infected. Humans can be infected with swine HEV via consumption of undercooked pork or through direct contact with infected pigs. To minimize the risk of zoonotic transmission, sensitive tools to assess the HEV infection status of pigs and pork products are needed. The objective of this study was to develop a fluorescent microbead-based immunoassay (FMIA) for the detection of IgG antibodies against swine HEV and compare it to an in-house enzyme-linked immunoassay (ELISA). Three sets of samples were utilized: (A) samples from pigs infected experimentally with different strains of HEV (positive controls, n = 72), (B) samples from known HEV-negative pigs (negative controls, n = 62) and (C) samples from pigs of unknown HEV infection status (n = 182). All samples were tested by both ELISA and FMIA. The results on the experimental samples with known HEV exposure indicate that both assays have a specificity of 100% while the sensitivity ranges from 84.6% (ELISA) to 92.3% (FMIA). The overall prevalence of HEV IgG antibodies in field samples from pigs with unknown HEV exposure was 21.9% (40/182) for the ELISA and 21.4% (39/182) for the FMIA. The two assays had an almost perfect overall agreement (Kappa = 0.92).