散发性急性戊型肝炎血清抗体动态变化和肝脏超微结构的病理观察

为探讨散发性戊型肝炎血清抗体动态变化，应用酶联免疫法（ＥＩＡ）检测了７例急性戊型肝炎患者抗戊型肝炎病毒（ＨＥＶ）ＩｇＧ和ＩｇＭ抗体，并对１例患者进行了肝超微结构病理检测。结果表明，发病后１０天至４５天内抗ＨＥＶ－ＩｇＧ和ＩｇＭ滴度最高，发病第４０天仍有肝细胞肿胀，胞浆空化和线粒体固缩等病理变化。患者血清抗－ＨＥＶＩｇＭ滴度在４５天后逐渐下降，２个月内全部消失，抗－ＨＥＶＩｇＧ消长情况与ＩｇＭ相似，但在７个月时仍有３例（４３％）抗体阳性。     

抗戊型肝炎病毒IgG的检测

应用ＥＬＩＳＡ方法对１９９０年沈阳一起暴发流行的戊型肝炎病人及对照人群７９份血清进行了抗戊型肝炎病毒（ＨＥＶ）ＩｇＧ的检测。显性感染病人与非疫区正常对照人群抗－ＨＥＶＩｇＧ检出率分别为５５．３２％（２６／４７）和２０．００％（２／１０），两者存在显著性差异（ｘ２＝４．１２，Ｐ＜０．０５）。显性感染病人急性期，病后８个月血清抗－ＨＥＶＩｇＧ检出率分别为１００．００％（８／８）和谐４１．９４（１３／３１），前者明显高于后者（ｘ２＝６．８３，Ｐ＜０．０１）。８例病人急性期和病后４个月血清抗体平均滴度分别为０．６８４和０．２３４，抗体滴度下降２．９２３倍，存在显著性差异（Ｔ＝２．２３９，Ｐ＜０．０５）。显性感染病人和ＳＧＰＴ升高无症状人群急性期血清间抗－ＨＥＶＩｇＧ检出率、抗体平均摘度存在明显差异（Ｐ＜０．０５，Ｔ＞１．９６０，Ｐ＜０．０５）。     

用重组结构区抗原检测抗丙型肝炎病毒IgM抗体方法的建立

目的检测抗丙型肝炎病毒（ＨＣＶ）结构区蛋白ＩｇＭ抗体。方法采用丙型肝炎病毒Ｃ，Ｅ１，Ｅ２区重组抗原混合包被和分别包被酶标板；用兔抗人γ链血清处理人血清标本，再用固相包被羊抗兔抗体吸附兔抗人γ链－人ＩｇＧ复合物，建立了抗－ＨＣＶＩｇＭ检测方法。结果对７６例丙型肝炎病人血清进行抗－ＨＣＶＩｇＭ检测，同时与逆转录－巢式聚合酶链反应（ＲＴ－ＰＣＲ）检测结果进行比较，两者具有相关性（Ｐ＜０００５）；ＩｇＭ抗体组成分析结果表明采用全片段Ｃ、Ｃ＋Ｅ２区抗原血清标本中的抗－ＨＣＶＩｇＭ检出率分别可达９６６％和１００％。结论ＨＣＶ的Ｃ、Ｅ２重组抗原用于抗－ＨＣＶＩｇＭ检测具有较高的敏感度和特异性。     

用重组结构区抗原检测抗丙型肝炎病毒IgM抗体方法？…

目的 检测抗丙型肝炎病毒（ＨＣＶ）结构区蛋白ＩｇＭ抗体。方法 采用丙型肝炎病毒Ｃ，Ｅ１、Ｅ２区重组抗原混合包被和分别包被酶标板；用兔抗人γ链血清处理人血清标本，再用固相包被羊抗兔抗体吸附兔抗人γ链－人ＩｇＧ复合物，建立了抗－ＨＣＶＩｇＭ检测方法。结果 对７６例现型肝炎病人血清进行抗－ＨＣＶ ＩｇＭ检测，同时与逆转录－巢式聚合酶链反应（ＲＴ＋ＰＣＲ）检测结果进行比较，再会得具有相关性（Ｐ〈０．００５     

散发性戊型肝炎病毒感染的诊断

用基因工程重组的戊型肝炎病毒基因结构区第二码框架和第二读码框架具有免疫表位的嵌合抗原，建立了间接酶联免疫法，检测散发性急性肝炎病人血清中抗－ＨＥＶＩｇＧ和ＩｇＭ抗体。在４６例急性肝炎病人中出抗－ＨＥＶＩｇＧ抗体阳性７例，阳性率为１５．２２％，７例ＩｇＧ抗体阳性中，有５例ＩｇＭ抗体也阳性，占７１．４％。     

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。     

用聚合酶链反应和放射免疫法检测肾移植术后人巨细胞病毒感染

将５０例肾移植术后发热病人的尿标本接种细胞，用聚合酶链反应（ＰＣＲ）技术检测第１代细胞培养上清中的人巨细胞病毒（ＨＣＭＶ）ＤＮＡ。结果１９例为ＨＣＭＶＤＮＡ阳性，阳性率为３８．００％，用固相放射免疫法（ＲＩＡ）检测ＨＣＭＶ抗原，阳性的有１５例，阳性率为２７．７７％。将５４倒尿标本接种人胎肺二倍体单层细胞，分离到６株ＨＣＭＶ。用ＥＬＩＳＡ检测６５例病人单份血清的ＨＣＭＶ－ＩｇＭ抗体，阳性的有１７例，检测４５例病人双份血清ＩｇＧ抗体，１１例的第２份血清的ＨＣＭＶ－ＩｇＧ抗体滴度比第１份有４倍以上升高。上述结果表明，ＰＣＲ检测结果与ＲＩＡ的检测结果基本符合；病人血清抗体的阳性率与ＰＣＲ检测的阳性率基本符合。     

巨细胞病毒感染与可溶性白细胞介素2受体的关系

应用酶联免疫吸附试验（ＥＬＩＳＡ）对１０４例育龄妇女的血清进行了巨细胞病毒（ＨＣＭＶ）ＩｇＧ、ＩｇＭ抗体的检测，同时用ＥＬＩＳＡ双抗体夹心法测定了不同感染状态下血清中可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）的水平，并将ｓＩＬ－２Ｒ水平与未感染ＨＣＭＶ的正常育龄妇女进行了比较。结果，育龄妇女中抗－ＨＣＭＶＩｇＧ的阳性率为８９．４％，ＩｇＭ的阳性率为９．６％，感染ＨＣＭＶ的妇女血清中ｓＩＬ－２Ｒ水平均大于未感染的对照组（１７８．１±５７．３Ｕ／ｍｌ），Ｐ＜０．０５，其中ＩｇＭ阳性者和ＩｇＭ、ＩｇＧ同时阳性者血清中ｓＩＬ－２Ｒ水平最高，分别为９１０±４６５．６Ｕ／ｍｌ和９０５±３４７．８Ｕ／ｍｌ，两者间的差异无显著性意义（Ｐ＞０．０５），但均大于仅抗－ＨＣＭＶＩｇＧ阳性者（４４６．８±１５８．９Ｕ／ｍｌ），Ｐ均＜０．０５。表明，ＨＣＭＶ感染可致ｓＩＬ－２Ｒ水平升高，并且活动性感染者上升明显。提示：ｓＩＬ－２Ｒ可能参与了ＨＣＭＶ的免疫致病机制。     

831名健康青年庚型肝炎病毒和人免疫缺陷病毒感染？…

目的 了解我国健康青年中庚型肝炎病毒（ＨＧＶ）和人免疫缺陷病毒的感染情况。方法 采用酶联免疫法（ＥＩＡ）检测６省８３１名健康青年血清中的ＨＧＶ和ＨＩＶ抗体，对抗－ＨＧＶＩｇＧ阳性的血清再用逆转录－巢式聚合酶链反应（ＲＴ－ＰＣＲ）检测ＨＧＶＲＮＡ。结果 发现抗－ＨＧＶ ＩｇＧ阳性率为２．５３％（２１／８３１），２１例阳性者中ＨＧＶ ＲＮＡ阳性８例，两者符合率为３８．１％；抗－ＨＩＶ均阴性。结论 我     

831名健康青年庚型肝炎病毒和人免疫缺陷病毒感染的调查

目的了解我国健康青年中庚型肝炎病毒（ＨＧＶ）和人免疫缺陷病毒的感染情况。方法采用酶联免疫法（ＥＩＡ）检测６省８３１名健康青年血清中的ＨＧＶ和ＨＩＶ抗体，对抗－ＨＧＶＩｇＧ阳性的血清再用逆转录－巢式聚合酶链反应（ＲＴ－ＰＣＲ）检测ＨＧＶＲＮＡ。结果发现抗－ＨＣＶＩｇＧ阳性率为２５３％（２１／８３１），２１例阳性者中ＨＧＶＲＮＡ阳性８例，两者符合率为３８１％；抗－ＨＩＶ均阴性。结论我国健康青年人群中确实存在ＨＧＶ感染。     

Diagnostic Performance of Five Assays for Anti-Hepatitis E Virus IgG and IgM in a Large Cohort Study

Determination of anti-hepatitis E virus (anti-HEV) antibodies is still enigmatic. There is no gold standard, and results obtained with different assays often diverge. Herein, five assays were compared for detection of anti-HEV IgM and IgG. Serum samples from 500 Swedish blood donors and 316 patients, of whom 136 had suspected HEV infection, were analyzed. Concordant results for IgM and IgG with all assays were obtained only for 71% and 70% of patients with suspected hepatitis E, respectively. The range of sensitivity for anti-HEV detection was broad (42% to 96%); this was reflected in the detection limit, which varied up to 19-fold for IgM and 17-fold for IgG between assays. HEV RNA was analyzed in all patients and in those blood donors reactive for anti-HEV in any assay, and it was found in 26 individuals. Among all of the assays, both anti-HEV IgG and IgM were detected in 10 of those individuals. Twelve had only IgG and, in 7 of those 12, IgG was only detected with the two most sensitive assays. Three of the HEV-RNA-positive samples were negative for anti-HEV IgM and IgG in all assays. With the two most sensitive assays, anti-HEV IgG was identified in 16% of the blood donor samples and in 66% of patients with suspected HEV infection. Because several HEV-RNA-positive samples had only anti-HEV IgG without anti-HEV IgM or lacked anti-HEV antibodies, analysis for HEV RNA may be warranted as a complement in the laboratory diagnosis of ongoing HEV infection.     

Hepatitis E virus seroprevalence in acute viral hepatitis in a developed country confirmed by a supplemental assay

Hepatitis E virus (HEV) infection is prevalent among cases of acute viral hepatitis in young adults in developing countries. HEV infection is not restricted to endemic areas, but would appear to be worldwide in distribution. In order to document the incidence of HEV infection in acute hepatitis cases in a developed country, IgG and IgM anti-HEV antibodies and HEV RNA were tested in 101 Caucasian patients with acute viral hepatitis; 92 of these cases had markers of acute viral hepatitis other than HEV. Forty-seven (46.5%) cases had IgG anti-HEV; IgM anti-HEV and HEV viremia were not detected. As the incidence of anti-HEV was higher than would be expected, the possibility of the occurrence of false positive results was subsequently investigated. Supplemental antibody testing, using a broadly reactive epitope region, reduced the frequency of anti-HEV to 17%. Therefore, supplemental antibody testing confirms the hepatitis E virus seroprevalence in a developed country. Since IgM anti-HEV and HEV viremia were not detected, persons with IgG anti-HEV may be “subclinical HEV cases,” or have long-lived antibodies in their circulation. © 1996 Wiley-Liss, Inc.     

IgM and IgG antibodies to hepatitis E virus (HEV) detected by an enzyme immunoassay based on an HEV-specific artificial recombinant mosaic protein

To develop an enzyme immunoassay (EIA) for IgM antibody to hepatitis E virus (HEV) (IgM anti-HEV) and IgG antibody to HEV (IgG anti-HEV), a synthetic gene encoding several liner immuno-dominant antigenic epitopes from HEV structural proteins was assembled as a chimeric recombinant mosaic protein (Mpr) with glutathione S-transferase and used as an immunodiagnostic target. In addition, a neutralization confirmation test was developed using individual synthetic peptides. Among 614 patients with acute hepatitis from 10 geographically distinct outbreaks, IgG anti-HEV was found in 546 (88.9%), with a range of 77–100% depending on the outbreak. Of 130 patients tested for IgM anti-HEV, 126 (96.9%) were positive. Among patients tested within 4 months of onset of jaundice, 37/37 (100%) were IgG anti-HEV positive. For patients from whom sera were collected 1–16 days after onset of jaundice, the geometric mean IgG titer (GMT) was 1:47,000; the GMT increased to 1:70,710 30–40 days after onset of jaundice and decreased to 1:1,778 3–4 months after the onset of jaundice. For patients tested 6–8 months after onset of jaundice, 11/12 (92%) were IgG anti-HEV positive, and the GMT was 1:2,908. IgM anti-HEV was detected in 43/43 (100%) sera collected 1–40 days after onset of jaundice, and the GMT for IgM anti-HEV was 1:10,000 at that time. For sera collected 3–4 and 6–12 months after onset of jaundice, 7/14 (50%) and 5/12 (40%) respectively, were IgM anti-HEV positive. In conclusion, an artificial mosaic protein composed of linear antigenic epitopes from open reading frame 2 (ORF2) and ORF3 of HEV has been successfully applied to the development of a sensitive and specific EIA for the detection of IgG and IgM anti-HEV activity. These assays were used for the verification of HEV infection in outbreak settings and for the diagnosis of HEV infection in sporadic cases. © 1996 Wiley-Liss, Inc.     

Simultaneous detection of immunoglobulin A (IgA) and IgM antibodies against hepatitis E virus (HEV) Is highly specific for diagnosis of acute HEV infection

Serum samples collected from 68 patients (age, mean +/- the standard deviation [SD], 56.3 +/- 12.8 years) at admission who were subsequently molecularly diagnosed as having hepatitis E and from 2,781 individuals who were assumed not to have been recently infected with hepatitis E virus (HEV; negative controls; 52.9 +/- 18.9 years), were tested for immunoglobulin M (IgM) and IgA classes of antibodies to HEV (anti-HEV) by in-house solid-phase enzyme immunoassay with recombinant open reading frame 2 protein expressed in the pupae of silkworm as the antigen probe. The 68 patients with hepatitis E had both anti-HEV IgM and anti-HEV IgA. Among the 2,781 controls, 16 (0.6%) had anti-HEV IgM alone and 4 (0.1%) had anti-HEV IgA alone: these IgA/IgM anti-HEV-positive individuals were not only negative for HEV RNA but lack IgG anti-HEV antibody as well (at least in most of the cases). Periodic serum samples obtained from 15 patients with hepatitis E were tested for HEV RNA, anti-HEV IgM, and anti-HEV IgA. Although HEV RNA was detectable in the serum until 7 to 40 (21.4 +/- 9.7) days after disease onset, both IgM and IgA anti-HEV antibodies were detectable until 37, 55, or 62 days after disease onset in three patients and up through the end of the observation period (50 to 144 days) in 12 patients. These results indicate that detection of anti-HEV IgA alone or along with anti-HEV IgM is useful for serological diagnosis of hepatitis E with increased specificity and longer duration of positivity than that by RNA detection.     

Incidence and seroprevalence of hepatitis E virus infection in pregnant women infected with hepatitis B virus and antibody placental transfer in infants

BackgroundHepatitis E has poor outcomes in pregnant women. Superinfection of hepatitis E virus (HEV) in patients infected with hepatitis B virus (HBV) may worsen liver disease.ObjectivesTo estimate the incidence and seroprevalence of HEV infection among HBV-infected pregnant women, to investigate the transplacental transfer of maternal anti-HEV IgG, and to compare the maternal and neonatal outcomes in anti-HEV positive and negative pregnant women.Study designTotally 391 HBV-infected pregnant women were recruited from April 2012 to October 2014. Paired mothers and infants were followed up at an average 9.8 months postpartum. Anti-HEV IgG and IgM were tested by ELISA.ResultsOf the pregnant women, none was anti-HEV IgM positive and 42 (10.7%) were IgG positive. At the follow-up, 3 seronegative women converted to anti-HEV IgG positive, with an estimated incidence of 17 per 1000 person-years. No significant differences of gestational age, preterm birth rate, Apgar score and birthweight were observed between newborns of anti-HEV IgG positive and negative mothers. Of the 42 neonates born to anti-HEV IgG positive mothers, 38 (90.5%) had anti-HEV IgG in their cord blood. The neonatal and maternal anti-HEV IgG levels were positively correlated (r = 0.827, p < 0.05). All infants were negative for both anti-HEV IgM and IgG at the follow-up.ConclusionsHBV-infected pregnant women rarely have novel HEV infection during late pregnancy in Jiangsu, China. Maternal anti-HEV IgG efficiently transfers into the fetuses, and disappears in infants before 10 months old.     

Detection of immunoglobulin M antibodies to hepatitis E virus by class capture enzyme immunoassay

The measurement of antibodies to hepatitis E virus (anti-HEV) has been essential for understanding the epidemiology of hepatitis E. Studies to determine the prevalence of HEV infections require a reliable serologic assay that is sensitive and specific. It is also important to distinguish the acute from the convalescent phase of an infection; this usually requires the detection of the immunoglobulin M (IgM) class of antibody. Few enzyme immunoassays (EIAs) that measure IgM anti-HEV have been described, and most have utilized the sandwich method. The present study describes an EIA that detects IgM anti-HEV by antibody class capture methodology. The assay was validated by using serum and/or plasma panels from experimentally infected nonhuman primates. It was used to demonstrate an anamnestic response and the reappearance of IgM anti-HEV in a chimpanzee experimentally challenged with HEV at two different times 45 months apart. The class capture method was more sensitive than the sandwich EIA when used to test clinical samples from two hepatitis E epidemics in Pakistan; it also had the advantage of distinguishing IgM anti-HEV in the presence of high titers of IgG anti-HEV.     

Serodiagnosis of La Crosse virus infections in humans by detection of immunoglobulin M class antibodies.

Sera from 92 humans with illnesses clinically compatible with those caused by California serogroup virus infections were tested for antibody to La Crosse (LAC) virus by using the immunoglobulin M (IgM) antibody-capture enzyme-linked immunosorbent assay (MAC ELISA), the IgG ELISA, and the hemagglutination inhibition (HI), complement fixation and serum dilution-plaque reduction neutralization tests. On the reported day of onset of illness in 18 individuals, 94% had IgM antibody, 50% had neutralization antibody, 33% had HI antibody, and 11% had IgG antibody. Neutralization, HI, and IgG antibody prevalence rates increased thereafter, whereas IgM antibody prevalence remained high (92% 2 or more weeks after the onset of illness). It was concluded that the MAC ELISA is a sensitive test for the presence of antibody to LAC virus. The sensitivity of the MAC ELISA and the rapidity with which it can be performed appear to provide a powerful tool for the clinically relevant serodiagnosis of LAC virus infections in humans.