应用捕获ELISA检测肾综合征出血热患者尿液中特异性IgM抗体

目的 检测紧综合征出血热患者尿液中特异性IgM抗体,研究早期诊断方法。方法：用MacELISA法检测不同病日HFRS病人尿液中特异性IgM抗体。结果：HFRS病人尿夜中特异性IgM抗体总阳性率为76.47%,第3病日即可出现阳性,7－9病日阳性率可达83.87%,与同期HFRS病人血清抗体检测对比无显著性差异（P＞0.05）。结论：用MacELISA检测HFRS尿液中特异性IgM抗体敏感性高,特异性强,适合早期诊断。     

免疫滴金技术检测肾综合征出血热特异性抗体458例

肾综合征出血热 (HFRS)是由汉坦病毒 (HV )引起的自然疫源性传染病。对该疾病的早期诊断是提高临床治愈率的关键。目前常用的IgM抗体酶联免疫吸附试验(MacELISA)检测特异性IgM抗体、间接免疫荧光法 (IFA)检测特异性IgG抗体和逆转录 聚合酶链反应 (RT PCR)法基因检测手段 ,在基层单位应用都受到条件和速度的限制 ,为了寻找一种更为简便、快速、特异、灵敏的检测方法 ,我们在近年开展了免疫滴金法检测IgM和IgG抗体研究的基础上 ,进一步优化条件 ,取得了较为满意的结果。材料与方法一、一般资料本组病例均为本院 1998年 1月 1日～…     

日本血吸虫病患者尿液中循环抗原和抗体联合检测的诊断价值

[目的 ]探讨尿液中血吸虫循环抗原和抗体检测对日本血吸虫病的诊断价值。 [方法 ]用单克隆抗体夹心 ELISA法检测日本血吸虫病患者尿液中循环抗原 ,间接ELISA检测尿液中特异性抗体。 [结果 ]10例急性血吸虫病和 6 1例慢性血吸虫病患者尿液中循环抗原的阳性率分别为 6 0 %和 40 % ,特异性抗体的阳性率分别为 80 %和 6 1 7%。两者联合检测的总阳性率分别为 10 0 %和 71 7%。 10 0例健康对照者尿液中仅 3%出现假阳性。 [结论 ]检测尿液中日本血吸虫循环抗原和特异性抗体简便、实用 ,为一种非损伤性的血吸虫病诊断方法。     

抗体捕获ELISA检测IgM抗体在新疆出血热病人早期诊断中的应用

采用抗人μ链抗体包被,捕获病人血清标本中的IgM抗体,再加已知的特异性病毒抗原与IgM抗体结合,用抗新疆出血热病毒单克隆抗体酶结合物与病毒抗原结合病人血清中IgM抗体的方法,测定急性期患者特异性IgM抗体,探讨其早期诊断的价值。对8例发病后不同时期的XHF患者进行检测的结果,发病第5天即可检出IgM抗体且滴度已达较高水平,于第14天达高峰后开始下降,但阳性可持续70天以上。而IgG抗体对照检测显示虽然在1/3急性期病人发病第6天即可检出,但滴度很低,第14日后迅速升高,至第70天仍然保持很高滴度。实验结果表明用本法检测IgM抗体进行早期诊断的意义可与病毒分离及特异性病毒抗原的检测结果相比。为XHF的早期诊断提供了一种特异性和敏感性高,并且稳定可靠的诊断工具。     

黄疸婴儿人巨细胞病毒的检测

目的: 探讨黄疸婴儿HCMV感染的诊断.方法:FQ-PCR检测248例黄疸婴儿和50例对照组患儿尿液中HCMV DNA, 化学发光免疫分析法(CLIA)检测黄疸婴儿血清HCMV IgM抗体, 在尿液HCMV DNA阳性黄疸婴儿中, 比较血清HCMV IgM抗体阳性与阴性组HCMV DNA拷贝数的差异.结果: 黄疸婴儿尿液HCMV DNA的阳性率为41.5%, 对照组患儿阳性率为2.0%, 两者有统计学意义(P<0.01). CLIA检测黄疸婴儿血清HCMV IgM抗体的阳性率为19.8%. 在尿液HCMV DNA检测阳性的黄疸患婴儿中, IgM抗体阳性组的尿液HCMV DNA拷贝数显著高于阴性组(t = 5.51, P<0.01).结论:FQ-PCR检测尿液HCMV DNA是早期诊断婴儿HCMV感染的敏感有效的方法.     

酶联免疫吸附试验检测梅毒患者血清中抗苍白螺旋体IgM抗体

目的评价抗苍白螺旋体IgM抗体检测对梅毒的临床意义.方法用酶联免疫吸附试验(ELISA)对72例梅毒患者检测了特异性IgM抗体,并与快速血浆反应素环状卡片试验(RPR)、梅毒螺旋体明胶颗粒凝集试验(TPPA)的检测结果进行比较分析.结果血清抗苍白螺旋体IgM抗体在一期梅毒的阳性率为73.3%(11/15),在二期梅毒的阳性率为88.9%(16/18),二者差异无显著的统计学意义(χ2＝1.6363,P＞0.10).在潜伏梅毒,IgM抗体阳性率为26.1%(6/23),非常显著地低于早期显性梅毒(χ2＝17.6189,P＜0.005).在一期、二期和潜伏梅毒,RPR和TPPA的阳性率均为100%.入组前2～24个月已经正规抗梅治疗的梅毒16例,其中IgM抗体阳性2例.结论 ELISA法检测特异性IgM抗体诊断一期梅毒并不优于RPR和TPPA.IgM抗体在潜伏梅毒敏感性低,其诊断应依靠RPR和TPPA.目前不推荐单独检测抗梅毒IgM抗体来监测病情和判断疗效.抗苍白螺旋体IgM抗体的临床意义有待更深入的研究.     

应用免疫酶斑点法检测流行性出血热病人血清IgM抗体的研究

<正> 目前流行性出血热(EHF)特异的实验诊断方法相继建立,但大多以检测EHP病人血清中IgG 抗体为多,有些方法由于要求实验条件较高,难以推广和普及。近年邢峥等应用 IgM抗体捕获ELISA法(MacELISA)进行 EHF的早期诊断获得满意的效果。最近我们在应用免疫酶斑点法(Immune Enzyme DotAssay,IEDA)检测 EHF 病人血清和尿中IgG 抗体的基础上,试用于 IgM的检测,取得一定的敏感性和特异性。现将结果报告如下。     

血吸虫感染小鼠早期诊断抗原的研究

目的对已知的6种日本血吸虫抗原的早期诊断价值进行评价,为研制用于哨鼠早期诊断的免疫试剂提供候选抗原。方法用日本血吸虫尾蚴感染小鼠,收集感染前及感染后不同时间的小鼠血清。采用重组日本血吸虫中国大陆株23kDa膜蛋白大亲水性肽段与谷胱甘肽的融合表达蛋白（GST-HD）、可溶性虫卵抗原（SEA）、血吸虫四跨膜蛋白第二亲水基团（TSP2HD）、血吸虫虫卵蛋白（IPSE）、日本血吸虫虫卵毛蚴抗原（SjMP-10）及日本血吸虫信号蛋白（Sj14-3-3）作为诊断抗原,采用酶联免疫吸附试验检测感染血吸虫后不同时间小鼠血清中特异性抗体IgM和IgG水平,通过分析感染后不同时间点抗原特异性抗体水平变化及阳性率,筛选具有血吸虫感染早期诊断价值的抗原分子。采用免疫印迹试验进一步验证其对血吸虫急性感染早期诊断的价值。结果感染后第18、21、28天,抗GST-HD抗体IgM阳性率分别为60%、70%、100%,特异性IgG阳性率分别为40%、60%、90%;抗SEA抗体IgM阳性率为50%、60%、90%,特异性IgG阳性率为20%、50%、70%;抗TSP2HD抗体IgM阳性率为30%、40%、50%,特异性IgG阳性率为20%、30%、70%;抗IPSE抗体IgM阳性率为20%、30%、50%,特异性IgG阳性率为20%、30%、60%;抗SjMP-10抗体IgM阳性率为10%、20%、20%,特异性IgG阳性率为10%、20%、30%;抗Sj14-3-3抗体IgM阳性率为0、10%、20%,特异性IgG阳性率为0、10%、30%。以GST-HD融合蛋白和SEA为抗原,检测小鼠早期感染血吸虫的敏感性高于Sj14-3-3、IPSE、TSP、MP-10,检测IgM的敏感性高于IgG。免疫印迹试验结果显示,SEA中分子量在73kDa左右的蛋白条带可被感染后1周小鼠血清所识别,并随着时间推移反应加强。GST-HD最早出现反应的血清是感染后第10天小鼠血清,反应强度在感染后第5周达到最强。结论重组GST-HD融合蛋白与SEA中分子量约73kDa的蛋白分子具有血吸虫感染早期诊断价值,免疫印迹试验的敏感性比酶联免疫吸附试验高。     

免疫酶染色试验检测斯氏狸殖吸虫抗体的研究

目的探索用免疫酶染色试验检测斯氏狸殖吸虫病抗体的敏感性和特异性。方法从病犬肺虫囊中收集斯氏狸殖吸虫成虫,冰冻切片,制作抗原片。采用免疫酶染色试验(IEST)检测斯氏狸殖吸虫病人和病鼠血清抗体。试验设健康大鼠血清对照及其他寄生虫病血清对照。结果斯氏狸殖吸虫病人和病鼠血清特异抗体阳性率分别为96.7%和97.4%。实验动物从感染后第2周开始抗体检测阳性,第4周阳性率达97.4%,持续10周。结论免疫酶染色试验敏感性高、特异性强,对斯氏狸殖吸虫病的早期诊断有重要参考价值。     

血清与脑脊液中病原抗体检测对病毒性脑炎的诊断价值

目的研究病毒性脑炎患者血清与脑脊液标本中病毒特异性抗体IgM阳性率的差异,并探讨其诊断意义。方法回顾同期进行了血清与脑脊液检测病毒特异性抗体IgM的89例病毒性脑炎住院患者资料,统计抗体阳性率,比较：（1）两种标本中病毒特异性抗体阳性率的差异;（2）一种标本检测与两种标本同时检测病毒特异性抗体阳性率的差异。结果（1）血清标本病毒特异性抗体的阳性率为53.93%,脑脊液标本病毒特异性抗体的阳性率为34.83%,两者比较有显著统计学差异（P〈0.025）。（2）两种标本同时检测病毒特异性抗体阳性率为71.91%,与任何一种标本检测的阳性率比较,有显著统计学差异（P〈0.02）。结论（1）检测血清中病毒特异性抗体可获得较脑脊液高的阳性率;（2）两种标本同时检测病毒特异性抗体是一种提高病毒性脑炎病原学诊断阳性率的有效方法。     

Hyper IgM syndromes

The immune defense against extracellular pathogens is largely dependent on antibody production. Class switch recombination and somatic hypermutation shape the secondary antibody repertoire in peripheral lymphoid tissue. In the past few years, a series of primary immune deficiencies characterized by defects in these processes and collectively referred to as hyper-IgM syndromes, have been described. Careful investigation of these rare "experiments of nature" has enabled to identify novel genes and molecular events that drive terminal B-cell differentiation. Abnormalities in these genes are likely involved also in lymphoid tumorigenesis and autoimmunity.     

The hyper IgM syndrome

The hyper IgM syndrome is a rare, inherited immune deficiency disorder resulting from defects in the CD40 ligand/ CD40-signaling pathway. X-linked hyper IgM is caused by defects in the CD40 ligand gene, while autosomal recessive hyper IgM is caused by defects in the CD40-activated RNAediting enzyme, activation-induced cytidine deaminase, which is required for immunoglobulin isotye switching and somatic hypermutation in B cells. The loss of interaction between CD40 and its ligand in X-linked hyper IgM results in an impairment of T cell function, of B cell differentiation, and of monocyte function, while only B cell differentiation appears to be affected in autosomal recessive hyper IgM. With genetic defects in the hyper IgM syndrome identified, it is possible to diagnose patients definitely, to perform genetic screening, and to delineate the clinical manifestations of this syndrome. Further research may lead to novel and definitive therapeutic options for patients with hyper IgM syndrome.     

In vitro synthesis of IgM and IgM rheumatoid factor in seronegative arthritides

Summary In vitro patterns of IgM and IgM rheumatoid factor (RF) synthesis exhibited by peripheral blood B cells (MNL) obtained from healthy individuals as well as patients with seropositive and seronegative rheumatoid arthritis (RA) have been previously examined. The present study was performed in a group of patients with non-rheumatoid seronegative arthritis (SNA) in order to compare patterns of in vitro IgM and IgM RF synthesis to that previously observed with seropositive and seronegative RA. Eighteen patients with SNA (4 ankylosing spondylitis, 10 psoriatic arthritis, and 4 unclassified variant disease) were studied as well as 18 healthy adult controls.Spontaneous release of IgM RF by MNL was not observed in SNA or controls. In contrast, IgM RF was detected in pokeweed mitogen (PWM)-stimulated MNL culture supernatants from 9/18 SNA (mean±SD=17.0±9.9 ng/10⁶ cells), and 10/18 normal controls (16.7±9.9 ng). Synthesis of IgM by PWM-stimulated MNL from SNA (2 062±1 200 ng) was significantly less than observed with MNL from controls (4 093±1 896 ng) (P<0.001). There were no differences among the various SNA subsets with regards to the levels of IgM and IgM RF produced either spontaneously or after PWM stimulation. IgM RF constituted a small fraction of the total IgM in SNA and normals (0.9% and 0.5%, respectively). This is clearly distinct from seropositive RA in which we have previously established that IgM RF constitutes a substantial fraction of the total IgM (10.7%) (P<0.01). IgM and IgM RF production did not appear to be significantly influenced by immunosuppressive medication.The results suggest that regulatory mechanisms governing the expression of RF are intact in patients with SNA and that lack of in vivo RF production in these patients cannot be related to an absolute deficiency of PWM-inducible mature RF committed B cells.     

Mechanism of IgM Polymerization

The stoichiometry of J chain in pentamer IgM has been determined by measuring the radiolabeled thiols in the constituent chains after complete reduction and alkylation of the polymer. One mole of J was found to be disulfide bonded to 1 mol of pentamer. The linkage of J chain in IgM has been determined by correlating the J disulfides cleaved with the subunits released after limited reduction and alkylation of the polymer. The analyses showed that: (a) Significant amounts of monomers, as well as small quantities of dimers, trimers, and tetramers, were generated by the reducing conditions employed. (b) The number of J disulfide bonds broken did not correspond to the extent of depolymerization. (c) No J disulfides were cleaved in the J-containing dimer products of the limited reduction. These data demonstrated that the J chain is located as a disulfide clasp between two of the IgM monomer subunits. From the observed linkage, the assembly of IgM is postulated to proceed by a series of sequential disulfide exchanges beginning with the formation of the J-containing dimer.     

IgM kappa gastric plasmacytoma

A case of gastric plasmacytoma in a 50-year old woman was reported. Immunofluorescent and immunoperoxidase studies were performed. Polyclonal antibodies reactive with immunoglobulin chains and a panel of 14 monoclonal antibodies reactive with B and T cells, and epithelial cells were used. These studies showed that the tumor cells produced IgM Kappa molecules whereas no monotypic immunoglobulin could be detected in the serum and urine. On the other hand the tumor cells had the immunologic phenotype of plasma cells. This helps diagnosis: some lymphomas with plasmocytic differentiation could also produce a monotypic immunoglobulin. Treatment using a combination of surgery, radio and polychimiotherapy was effective, leading to complete remission.