人疱疹病毒6型感染与白血病关系的初步研究

白血病是来源于造血系统的一种恶性肿瘤，好发于儿童和青少年，其病因和发病机制尚不完全清楚，但病毒病因学较受重视。人疱疹病毒6型(HHV-6)是1986年发现的疱疹病毒科中的一个新成员。研究发现，HHV-6的感染与淋巴瘤的发生有一定关系，在白血病患者中具有较高的感染率。为了探讨HHV-6感染与白血病的关系，我们采用间接免疫荧光试验(IFA)和PCR，分别检测了白血病患者血清抗HHV-6 IgG和外周血单个核细胞(PBMC)中HHV-6 DNA的序列。     

人疱疹病毒6型感染与淋巴瘤关系的初步研究

目的:研究人疱疹病毒6型(HHV-6)感染与淋巴瘤的关系。方法:用间接免疫荧光法及PCR,分别检测淋巴瘤患者和对照组患者血清中抗HHV-6 IgG及外周血单个核细胞(PBMC)中HHV-6 DNA序列。用免疫组化染色法,检测淋巴瘤患者淋巴结组织标本中HHV-6抗原。结果:淋巴瘤患者血清抗HHV-6抗体的阳性率为95.5％,几何平均滴度为1:123,PBMC中HHV-6 DNA的检出率为59.1％,均明显高于对照组(P<0.05)。免疫组化染色检测的5例淋巴瘤患者淋巴结组织标本中,4例HHV-6抗原阳性。结论:HHV-6感染可能与淋巴瘤的发病有关。     

口腔鳞癌患者淋巴细胞对人疱疹病毒6型增殖应答受损

目的研究口腔鳞癌患者淋巴细胞对人疱疹病毒6型（HHV-6）特异性增殖应答,初步探讨HHV-6在口腔鳞癌发病机制中的作用。方法间接免疫荧光法（IIF）检测血浆中抗HHV-6IgG;免疫微磁珠分离CD4^＋T及CD8^＋T细胞;3H-TdR法检测CD4^＋T和CD8^＋T细胞及PBMCs的增殖水平;FACS分析CD4^＋C25^＋调节性T细胞（Treg）的比例。结果口腔鳞癌组血浆中抗HHV-6IgG阳性数为8/8,正常对照组为12例（12/20）;口腔鳞癌组PBMCs及CD4^＋T细胞对HHV-6的增殖水平显著低于HHV-6潜伏感染组与未感染组（P〈0.05）;HHV-6潜伏感染组PBMCs及CD4^＋T细胞对HHV-6的增殖水平显著低于未感染组（P〈0.05）;口腔鳞癌组外周血中CD4^＋C25^＋Treg比例明显高于HHV-6潜伏感染组与未感染组（P〈0.05）。结论口腔鳞癌患者的HHV-6特异性CD4＋T细胞增殖应答减弱,可能在口腔鳞癌的发生发展中起一定作用。     

冠状病毒感染细胞抗原片法检测SARS患者血清中特异性抗体

目的：检测SAILS患者血清中特异性抗冠状病毒抗体及其滴度变化，协助临床确诊SAILS。方法：用本次爆发SAILS患者鼻腔抽吸液分离的冠状病毒感染新生恒河猴肾细胞，制备抗原片，用间接免疫荧光法检测患者体内急性期和恢复期血清中的抗冠状病毒抗体。结果：汕头地区7例SAILS患者血清内均有特异性抗冠状病毒抗体，且恢复期抗体滴度比急性期的滴度增高4倍以上。结论：用冠状病毒感染细胞抗原片对怀疑感染SAILS的患者血清进行特异性抗体检测，可协助临床确定SAPS的诊断。     

抗MPO抗体的ELISA检测和应用

本文应用Ｍａｔｈｅｓｏｎ方法从人白细胞中提取髓过氧化物酶ＭＰＯ，建立ＥＬＩＳＡ法检测６３份血清标本中抗ＭＰＯ抗体，同时与间接免疫荧光法（ＩＩＦ）作比较，结果是：１９例结节性多动脉炎（ＰＡＮ）患者中６例抗ＭＰＯ抗体阳性（３２％），较ＩＩＦ法阳性率提高（１９例中１例），而２２例正常人及２２例其他肾脏病患者血清均为阴性。提示ＥＬＩＳＡ法既可从分子水平鉴定系统性血管炎致病的特异性抗体，又是系统性血管炎敏感的诊断方法。     

检测抗人疱疹病毒6型IgG的间接免疫荧光试验的建立及其应用

目的 :建立检测血清中人疱疹病毒 6型 (HHV 6 )IgG的间接免疫荧光试验 (IFA)。方法 :用HHV 6国内分离株感染人脐带血单个核细胞制备抗原片 ,建立检测HHV 6IgG的IFA方法 ,并用于育龄期妇女血清流行病学调查。结果 :建立的IFA具有特异性。对 116份育龄期妇女血清标本检测表明 ,HHV 6IgG的阳性率为 72 .4 % ,几何平均滴度 (GMT)为 1∶6 1;在孕妇和正常未孕妇女之间 ,以及不同孕期的孕妇之间 ,HHV 6IgG的阳性率和GMT均无差异 (P >0 .0 5 )。结论 :建立了具有特异性的IFA法 ,可用于对育龄妇女HHV 6感染率的流行病学调查     

汉滩病毒S基因真核表达载体的构建及免疫小鼠的初？…

目的 研究汉滩病毒Ｓ片段编码区基因免疫小鼠的作用。方法 利用基因重组技术，构建含汉滩病毒Ｓ片段编码区基因的真核表达质粒。用此质粒直接注射到ＢＡＬＢ／ｃ小鼠骨骼肌内进行ＤＮＡ免疫，用间接免疫荧光法检测免疫小鼠血清中汉滩病毒抗体。结果 在初次免疫的小鼠血清中检测出有低滴度的汉滩病毒抗体。加强免疫后，抗体水平显著上升，免疫荧光抗体（ＩＦＡＴ）滴度最高达１：６４０。约８７％的免疫动物发生了血清抗体阳转。抗     

EHF病毒感染家兔后血清特异性抗体水平的动态观察

本文报告了流行性出血热病毒(Epidemic haemorrhagic fever virus,EHFV)J10、A9株分别感染实验家兔后,用反向间接ELISA(RIELISA)和间接免疫荧光(IFA)方法,动态观察了家兔特异性IgG抗体水平的变化。发现特异性IgG于感染后第1周开始出现,以后抗体水平稳步上升,于第5—7周便达到最高水平,高滴度的抗体可以维持较长时间。结果还表明;RIELISA检测家兔抗体的敏感性明显优于IFA法。用IFA进行的交叉反应还证实;EHFV-J10、A9两种毒株的抗血清能与不同来源的数株病毒发生特异性结合反应。     

广州市2009年肾综合征出血热监测分析

目的分析广州市肾综合征出血热（HFRS）的流行特征和规律,探讨防制对策。方法采用人间疫情监测、健康人群抗体水平监测、宿主动物监测方法,其中实验室用间接免疫荧光法检测血清特异性IgG抗体,直接免疫荧光法检测鼠肺汉坦病毒（HV）抗原。结果共报告HFRS71例,死亡1例,发病率为0.69/10万,病死率为1.41%;排查病例抗体阳性率为16.14%（188/1165）,健康人群抗体阳性率为0.50%（2/400）;鼠密度为4.60%（361/7856）,鼠肺抗原阳性率为8.31%（30/361）,鼠血清抗体阳性率为7.43%（26/350）,优势鼠种为褐家鼠。结论广州市HFRS疫情形势较为平稳,但仍然存在病例数上升的压力,应加强疫情监测,落实防鼠灭鼠为主的综合性预防控制措施。     

狂犬病疫苗免疫后血清抗体水平的调查

狂犬病疫苗免疫后血清抗体水平的调查朱宝根，张晓娟，赵娟，傅晓琰我们用间接免疫荧光法（ＩＦＡ）检测了人血清狂犬病病毒ＩｇＧ抗体，分析了不同性别、年龄对狂犬病疫苗免疫效果的影响，并对免疫失败的原因进行了初步探讨。材料与方法１．标本来源暴露者狂犬病疫苗全程...     

Comparison of specific serological assays for diagnosing human herpesvirus 6 infection after liver transplantation

Cross-reactivity between human herpesvirus 6 (HHV-6) and human herpesvirus 7 (HHV-7) antibodies and the reliability of specific serological assays were analyzed for 12 patients with concurrent HHV-6 and HHV-7 antibody responses after transplantation with a liver from a living relative by using an immunofluorescence assay (IFA). A neutralizing antibody titer assay (NT) and an immunoblot assay (IB) designed to detect immunoglobulin M (IgM) antibody to the HHV-6 immunodominant 101-kDa protein were compared in the diagnosis of an active HHV-6 infection. A total of 9 of 12 patients demonstrated concurrent HHV-6 and HHV-7 antibody responses, including increased IgG titers and/or the presence of IgM by IFA, and were thus analyzed for cross-reactive antibody to heterologous virus. The average percentages of residual antibody to HHV-6 and HHV-7 after absorption with HHV-6 antigen were 32.6% (range, 6 to 50%) and 55.6% (range, 35 to 100%), respectively. All 12 patients were subsequently analyzed for HHV-6 antibody by using IB and NT. IB detected IgM antibody to the 101-kDa protein in 75% (9 of 12) of the recipients. A significant rise in the NT antibody titer was detected in the same nine samples. However, HHV-6 DNA was detected by PCR in only five of nine plasma samples collected from recipients with a specific serologic response against HHV-6.     

Comparison of Specific Serological Assays for Diagnosing Human Herpesvirus 6 Infection after Liver Transplantation

Cross-reactivity between human herpesvirus 6 (HHV-6) and human herpesvirus 7 (HHV-7) antibodies and the reliability of specific serological assays were analyzed for 12 patients with concurrent HHV-6 and HHV-7 antibody responses after transplantation with a liver from a living relative by using an immunofluorescence assay (IFA). A neutralizing antibody titer assay (NT) and an immunoblot assay (IB) designed to detect immunoglobulin M (IgM) antibody to the HHV-6 immunodominant 101-kDa protein were compared in the diagnosis of an active HHV-6 infection. A total of 9 of 12 patients demonstrated concurrent HHV-6 and HHV-7 antibody responses, including increased IgG titers and/or the presence of IgM by IFA, and were thus analyzed for cross-reactive antibody to heterologous virus. The average percentages of residual antibody to HHV-6 and HHV-7 after absorption with HHV-6 antigen were 32.6% (range, 6 to 50%) and 55.6% (range, 35 to 100%), respectively. All 12 patients were subsequently analyzed for HHV-6 antibody by using IB and NT. IB detected IgM antibody to the 101-kDa protein in 75% (9 of 12) of the recipients. A significant rise in the NT antibody titer was detected in the same nine samples. However, HHV-6 DNA was detected by PCR in only five of nine plasma samples collected from recipients with a specific serologic response against HHV-6.     

Evaluation of immunoassays for detection of antibodies to human herpesvirus 7.

An enzyme immunoassay (EIA), an immunoblot assay (IB), and an indirect immunofluorescence assay were developed for detection of human herpesvirus 7 (HHV-7) antibodies in human serum. Cross-absorption studies with EIA or IFA using HHV-7 and human herpesvirus 6 (HHV-6) antigens indicated that most human sera contain cross-reactive HHV-6 and HHV-7 antibodies and that the degree of cross-reactivity varies between individual serum specimens. Inhibition of homologous antibody activity by absorption with heterologous virus ranged from 0 to 57% by EIA. However, for every sample tested, absorption with homologous virus removed more activity than did heterologous virus. An 89-kDa protein was identified as an HHV-7-specific serologic marker by IB. Activity to this protein was not removed by absorption with HHV-6 antigen. Of the three assays, the EIA was the most sensitive (94%), while the IB was the most specific (94%). Approximately 80% of specimens collected from German adults and children older than 2 years were positive for HHV-7 antibodies by these assays.     

Enzyme-linked immunosorbent assay for detection of antibody to human herpesvirus 6.

The results obtained with an enzyme-linked immunosorbent assay (ELISA) for detection of human herpesvirus 6 (HHV-6) immunoglobulin G using a single 1:100 dilution of serum correlated well with those found by an indirect fluorescence microscopic assay (IFA) (r = 0.71). Concordant results were found in all 7 paired serum samples obtained from patients with acute primary infections and in 37 of 41 (90.24%) single serum samples. Fourteen serum samples (25%) which yielded nonspecific results by IFA were evaluable by ELISA. In a serologic survey using the ELISA, a disproportionate number of 12-month-old infants had low difference-of-optical-density values, suggesting that maternal antibody might persist beyond a year of age. This finding and the rises in antibody to HHV-6 found in patients with primary cytomegalovirus infections might lead to overestimation of HHV-6 infection rates in young children in seroprevalence studies. Other herpesvirus infections produced lesser effects on anti-HHV-6.     

Enzyme-linked immunosorbent assay for detection of IgG antibody to human herpesvirus 6

A lysate of human herpesvirus 6 (HHV-6)-infected cord blood mononuclear cells was used as antigen for enzyme-linked immunosorbent assay for the detection of IgG antibody to HHV-6. Antibody responses after exanthem subitum were well correlated with clinical recovery from the disease and the level of antibody activities was well correlated with indirect immunofluorescence assay and the neutralization test. Seroconversion to other human herpesvirus, including cytomegalovirus, was not observed in infants with exanthem subitum. All of the infants had by the age of 1 month antibodies to HHV-6, which decreased with age to the lowest level at the age of 3 to 6 months and then increased and reached the maximum level by 1 to 2 years of age. After 3 years of age, the prevalence was almost stable.     

Serological response of paediatric oncology patients to human herpesvirus-6

The serological response of paediatric oncology patients to human herpesvirus-6 (HHV-6) was investigated at presentation and during treatment. Sera from 66 patients presenting with malignancy and 66 controls were examined for anti-HHV-6 IgG by indirect immunofluorescence test (IFA) and enzyme linked immunosorbent assay (ELISA), and for anti-HHV-6 IgM by IFA. Serial samples from 45 children on chemotherapy were examined for anti-HHV-6 IgG by ELISA and sera from selected patients on chemotherapy were examined for IgM by IFA. The response of these patients to four other herpesviruses was also investigated. Ninety percent of presenting patients and controls were IgG positive for HHV-6 by IFA and ELISA. Anti-HHV-6 IgG as measured by the HHV-6 ELISA index declined overtime in patients on chemotherapy. Two presenting controls and one leukaemic child with a primary cytomegalo-virus seroconversion were anti-HHV-6 IgM positive. In the patient group seropositivity to herpesviruses (types 1–6) increased with age, the mean age of those with IgG to HHV-6 alone was 3.7 years compared to 6.8 years for those with antibodies to all five viruses. At the time of presentation paediatric oncology patients have a similar serological response to HHV-6 as age-matched controls and this IgG response wanes with treatment. Whether this is significant in terms of viral pathogenicity is not known and will require investigation of viral activity in these patients. © 1994 Wiley-Liss, Inc.     

Isotype immune response of IgG antibodies at the persistence and reactivation stages of human herpes virus 6 infection.

BACKGROUND: Infections with human herpes virus 6 (HHV-6) are very common. After primary infection, the virus remains latent and persists at low level in cells and tissues. Not usually associated with disease in the immunocompetent host, HHV-6 infection is a major cause of opportunistic viral infections in the immunosuppressed. The different stages of HHV-6 infection are difficult to characterize in the laboratory. OBJECTIVES: The aim of this paper was to assess the isotype patterns of IgG antibodies against HHV-6 in seropositive subjects during different stages of the virus activity. STUDY DESIGN: From a total of 190 human serum samples from 43 healthy children, 24 pregnant women and 24 patients with bone marrow transplants, 111 sera were processed by indirect immunofluorescence assay for the detection of IgG1, IgG2, IgG3 and IgG4 specific antibodies. The mean geometrical title (MGT) of the antibodies was calculated. RESULTS: All pregnant women had IgG1 (24/24; 100%; MGT 46). A 95% (41/43) of healthy infants had IgG1 (MGT 57). In bone marrow transplants, 58% (14/24) of the patients showed seroconversion (MGT 529) with an isotype response of IgG1 and IgG4 during the observation period. Remaining bone marrow transplant patients, who had the IgG without any variations (MGT 184), had isotype IgG1. CONCLUSIONS: These results revealed two different immune isotype response patterns. One of them is restrictive to IgG1 in the latent phase of HHV-6 infection in healthy children, pregnant women and transplant patients with stable levels of antibodies whereas IgG1 and IgG4 are detected in the reactivation of HHV-6 in transplant patients. The IgG isotype immune responses may contribute to the existing set of serological markers in characterizing the different stages of natural infection of HHV-6.     

Evaluation of the specificity and sensitivity of indirect immunofluorescence tests for IgG to human herpesviruses-6 and -7

Sera from 118 children aged up to 4 years were tested by indirect immunofluorescence for human herpesvirus-6 and -7 (HHV-6 and HHV-7) antibodies. Antibody results were confirmed as true positives if the relevant viral DNA was detected in saliva or, in some cases of primary infection, by the finding of the relevant DNA in cerebrospinal fluid or serum. Results from samples taken from the 15 children less than 6 months old showed that HHV-6 and/or HHV-7 antibody was either absent or present at low titre suggesting persistent maternal antibody rather than true infection. The sensitivity, specificity, positive and negative predictive values of the HHV-6 IgG test were therefore based on the data from the 103 children older than 6 months and the results were 95, 84, 91 and 90%, respectively. Likewise, the sensitivity, specificity, positive and negative predictive values of the HHV-7 IgG test were 95, 76, 84 and 93%, respectively. There was limited cross-reactivity between HHV-6 and HHV-7 antibodies; where both HHV-6 and HHV-7 antibodies are detected, titres above 32 may be accepted as true positives but lower titres require confirmation by detection of the relevant viral DNA or, in the case of primary infection, by a rising antibody titre.     

An enzyme-linked immunosorbent assay for IgG antibodies to human herpes virus 6

An indirect enzyme-linked immunosorbent assay (ELISA) with human herpes virus 6 (HHV6) membrane antigen was compared with indirect immunofluorescence assay (IFA) for measurement of HHV6 IgG antibodies. Five hundred serum samples from 403 Swedish patients with suspected symptomatic Epstein-Barr virus (EBV) infections were examined. The specificity of the ELISA compared with IFA was 98.7% and the sensitivity was 98.4%. In 90% of the patients, IgG antibodies to HHV6 were detected with both assays. The highest HHV6 IgG titers were found mainly in patients with EBV or CMV infections, but HHV6 mononucleosis was not diagnosed. The same HHV6 antigen was assessed for IgM ELISA but was found to be of limited value due to high IgM reactivity with the control antigen. The HHV6 IgM ELISA requires further investigation. The IgG ELISA described is a reliable alternative to IFA for measurement of HHV6 IgG antibodies and for large scale epidemiological studies.