免疫斑点法检测抗精抗体的建立及其与ELISA法比较

目的建立检测抗精抗体的免疫斑点法,并与ELISA法进行比较。方法收集20份正常精液,混合,洗涤,混悬于0．5％NP-40 Tris—HCI缓冲液中,制备精予膜抗原,辣根过氧化物酶标记SPA,制备HRP—SPA,建立免疫斑点法检测抗精抗体,并对两种方法检测结果进行比较。结果免疫斑点法检测正常生育组30对夫妇共60例,其中阳性者1例为男性,阳性率为3．3％;检测不孕夫妇116对,共232例,阳性患者51例,阳性率为22．0％,其中男方占23例,阳性率为19．8％,女方占28例,阳性率为24．1％;检测习惯性流产夫妇53对,共106例,阳性患者16例,阳性率为15．1％,其中男方占2例,阳性率为3．8％,女方占14例,阳性率为26．4％。两种方法检测结果一致或差异不显著。结论免疫斑点法是一种比较稳定和敏感的方法,重复性、特异性好,结果易于判断,可应用于临床检验。     

国产微柱凝胶免疫检测试剂卡检测孕妇IgG血型抗体效价的实验研究

目的：比较微柱凝胶抗人球蛋白试验和传统试管抗人球蛋白试验检测O型孕妇血清中IgG抗-A和抗-B效价的差异,对国产微柱凝胶试剂卡检测孕妇IgG效价的决定值进行界定。方法：用国产微柱凝胶试剂卡和传统试管法平行检测524例丈夫为A型或B型的O型孕妇血清IgG抗-A和抗-B水平,经配对t检验、χ^2检验和回归分析,比较两种方法检测结果的差异并界定前者的决定值。结果：国产微柱凝胶试剂卡和传统试管法检测抗-A平均效价分别为249.98和120.85,抗-B为156.98和76.38,前一方法分别是后者2.07、2.06倍,差异均有显著性意义（t=19.64,P〈0.001;t=15.39,P〈0.001）;相关性分析得出两种方法测抗-A、抗-B效价均有高度的相关性,相关系数r分别是0.8857（t=30.76,P〈0.001）、0.8996（t=33.22,P〈0.001）,通过回归方程得出国产微柱凝胶试剂卡检测抗-A、抗-B效价的决定值分别为128、116;以IgG抗体效价≥64判为阳性,国产微柱凝胶试剂卡测抗-A、抗-B阳性率分别为73.66、52.29,明显高于传统试管法的47.33、34.35,均具有统计学意义（χ^2=69.00,P〈0.01;χ^2=47.00,P〈0.01）。结论：国产微柱凝胶试剂卡比传统试管法操作更简便、快捷,应用于孕妇产前IgG抗-A、抗-B效价检测比后者具有更高的灵敏度,对其决定值的界定,将为临床提供更可靠的诊断依据。     

两种检测方法测定抗甲状腺球蛋白抗体和抗甲状腺过氧化物酶抗体结果的比较

目的 应用化学发光免疫分析法（CLIA）和放射免疫分析法（RIA）分别测定血清抗甲状腺球蛋白抗体（抗TgAb）和抗甲状腺过氧化物酶抗体（抗TPOAb）.方法 采用CLIA法和RIA法测定304例不同类型甲状腺疾病患者和38名健康对照者血清抗TgAb和抗TPOAb水平.结果 CLIA法测定血清抗TgAb和抗TPOAb水平与RIA法测定的结果具有良好的一致性.结论 CLIA法测定血清抗TgAb和抗TPOAb可以用来替代RIA法.     

酶联法和免疫印迹法检测血抗卵巢抗体的比较

为比较酶联法（ELISA）和免疫印迹法（IBT）检测人抗卵巢抗体（AOA）的灵敏度和准确性,本文以正常妇女为对照,检测不明原因性不孕症和卵巢早衰（POF）患者血清AOA。结果表明POF组AOA阳性率分别为40％（ELISA）和80％（IBT）,不孕症组AOA阳性率为20％（ELISA）和60％（IBT）。两种方法比较,同组阳性率有显著差异（P<0.01）。IBT方法可区别非致病性和致病性AOA,提高诊断的准确性、实验表明ELISA和IBT均可用于检测AOA以了解卵巢功能降低之免疫性病因,但IBT方法更灵敏,更准确。     

血型不规则抗体的筛查和抗体特异性分析

目的确证微柱凝胶法血型不规则抗体筛检阳性患者的抗体特异性。方法应用微柱凝胶法对患者血浆(血清)进行血型不规则抗体筛查,对抗体筛查阳性患者的血标本再用凝聚胺法及间接抗人球蛋白试验进行抗体特异性鉴定和效价测定。结果在微柱凝胶法血型不规则抗体筛查阳性210例中,由非特异性抗体引起假阳性35例(16.7%),由血型不规则抗体引起阳性175例(83.3%)。在175例血型不规则抗体阳性患者中,Rh血型系统抗体156例(89.1%),MNSs血型系统抗体10例(5.7%),Lewis血型系统抗体5例(2.9%),Kidd血型系统抗体3例(1.7%),其中Rh血型系统合并Kidd血型系统抗体1例。结论对孕妇产前及受血者进行血型不规则抗体筛查、特异性鉴定及其效价测定,早期进行预防及选择不含相应抗原的血液输注,是防治新生儿溶血病和确保输血安全和有效的重要措施。     

抗Ce抗体及直接抗人球蛋白试验阳性血清学检测分析

目的 分析患者血清存在抗Ce抗体及直接抗人球蛋白试验(DAT)阳性的血清学检测结果及其配血对策。方法 采用微柱凝胶法(MGT)对患者进行血型鉴定及抗体筛选,对抗体筛选阳性患者血标本采用试管盐水法(NS)和间接抗人球蛋白试验(IAT)进行抗体特异性鉴定、吸收放散试验、抗体性质及抗体效价测定。采用NS、 MGT及IAT对患者与供血者血液进行交叉配血试验。结果 在21例患者中ABO血型A型6例、 B型9例、 O型4例、 AB型2例,Rh血型均为ccDEE,抗体筛选阳性,交叉配血不合。采用谱细胞进行抗体特异性鉴定,确认21例患者血清存在抗Ce抗体,抗体效价1∶4～1∶8,其中6例伴DAT阳性。结论 抗Ce抗体为免疫性IgG型抗体,可引起抗体筛选阳性、交叉配血不合、溶血性输血反应、新生儿溶血病及溶血性贫血。     

三种初筛方法检测HIV抗体的结果比较及评估

目的比较酶联免疫吸附试验（ELISA）、明胶颗粒凝集试验（PA）、胶体硒免疫层析法（ICA）检测人类免疫缺陷病毒抗体的结果，了解这三种方法的优劣，探讨临床实验室初筛诊断HIV方法的灵敏性和准确性。方法用卫生部HIV质控血清和免疫印迹法（WB）确认的HIV抗体阳性标本和阴性标本。分别用ELISA法、PA法及ICA法检测；用免疫印迹法确认的HIV阳性的标本作倍比稀释后，再分别用PA法、ELISA法及ICA法检测。结果80例经免疫印迹法确认的HIV阳性患者，ELISA法、PA法、ICA法的检出符合率分别为100％、98．8％、97．5％；经稀释法检测。ELISA法较PA法和ICA法敏感。其检测的准确性分别为97．5％、97．5％、92．5％。结论筛查HIV抗体方法最好选用ELISA法或PA法。以防漏检。     

甲状腺自身抗体的两种方法检测结果的比较及临床价值的探讨

目的对比两种方法检测甲状腺球蛋白抗体(TGA)、甲状腺微粒体抗体(TMA)、甲状腺过氧化酶抗体(TPoA)结果及其临床价值.方法对7例桥本氏病(HD)和甲状腺功能亢进40例,采用RIA法检测TGA、TMA及 CLIA法测定其TGA、TPoA的水平.结果 HD患者RIA法检测TGA、TMA和 CL IA法检测TGA、TPoA均高于正常水平,21例甲亢患者两种方法的检测结果均有1项以上增高;7例CLIA法检测TGA、TPoA有1项以上增高,但RIA法检测TGA、TMA均为阴性;其余11例两种方法测定结果均为正常水平.相关系数比较:方法内TGA 和TMA r=0.94(RIA法),TGA和TPoA r=0.15(CLIA法);RIA法和CLIA法TGA r=0. 55,TMA和TPoA r=0.63.结论①对于HD,TGA和TMA或TGA和TPoA 的同时检测;②对于甲亢CLIA法检测TGA和TPoA的阳性率明显优于RIA法检测TGA和TMA. 采用CLIA法直接测定TPoA更有临床价值.     

Rh血型抗C、抗D混合抗体的血清学检测及临床意义

目的 分析患者血清中同时含有Rh血型抗C、抗D混合抗体的血清学检测结果及临床意义.方法 采用卡式法对患者进行血型鉴定,对有疑问的患者血标本采用盐水法、凝聚胺法及间接抗人球蛋白试验进行不规则抗体筛选,对抗体筛选阳性的血标本进行抗体特异性鉴定.然后,对患者与供血者血液进行交叉配血试验.结果 患者1血型为O型,Rh血型为cc...     

四种方法检测抗线粒体抗体M2亚型性能比较

目的 探讨抗线粒体抗体M2亚型不同检测方法的分析性能、相关性及其对原发性胆汁性胆管炎的诊断价值.方法 选择2010年至2019年于北京同仁医院就诊的原发性胆汁性胆管炎(PBC)患者106例作为PBC组,同期其他自身免疫病组患者120例,100例体检健康人作为正常对照组.用免疫印迹、间接免疫荧光、酶联免疫吸附试验和胶体金...     

Direct correlation between human herpesvirus-8 seroprevalence and classic Kaposi's sarcoma incidence in Northern Sardinia

The human herpesvirus-8 (HHV-8) has been associated with the development of Kaposi's sarcoma. A high incidence of classic Kaposi's sarcoma has been described in Sardinia, an island West of Italy's mainland. Different seroepidemiological analyses have reported that prevalence of HHV-8 infection varies worldwide: a high HHV-8 seroprevalence has been shown in Italy. The present survey was carried out to evaluate the correlation between HHV-8 infection and classic Kaposi's sarcoma incidence in northern Sardinia. Blood samples were collected from 226 healthy donors born and resident in five different areas of North Sardinia. Seroprevalence to HHV-8 was determined searching antibodies to viral lytic proteins by immunofluorescence in sera diluted at 1:10. Classic Kaposi's sarcoma incidence data spanning a period of 23 years were examined in the areas studied. The present screening revealed that seroprevalence was 35%, within a range of 15.3-46.3% in the five areas, although it should be considered that the seroprevalence to HHV-8 can be established more accurately by the combined use of different assays. Age emerged as an important risk factor. Indeed, subjects aged > 50 years showed a higher seroprevalence to HHV-8 as compared with younger individuals. A strong direct correlation between HHV-8 prevalence and classic Kaposi's sarcoma incidence has been also observed. The wide diffusion of HHV-8 in Sardinia appears to represent an important factor in the high incidence of classic Kaposi's sarcoma reported in the island. However, additional co-factors, such as age, sex, genetic traits, or viral strain pathogenicity, are likely to play a role in the development of the disease.     

Antitumor Activity of Glycosylated Molluscan Hemocyanins via Guerin Ascites Tumor

As observed in most molluscan hemocyanins, high-mannose type glycans were identified in hemocyanins from Rapana venosa (RvH), Helix lucorum (HlH) and keyhole limpet (Megatura crenulata). In addition, a glycan with a branching structure containing xylose, fucose and terminal methyl hexose was identified in β-HlH. We have examined the immuno-adjuvant properties of hemocyanins, their derivatives and conjugates associated with the cell mediated immunity in experimental tumor-bearing animals with ascites tumor of Guerin. After immunization of the animals with the experimental vaccine preparations, the highest values of splenic lymphocytes were observed in groups immunized with the conjugates RvH-TAg, β-HlH-TAg and KLH-TAg (42.3%; 40.8% and 40.58%, respectively) than with the native hemocyanins (36.5%; 35.1% and 32.4%, respectively). The immunization of rats with the hemocyanins β-HlH, RvH and KLH and their conjugates, prolonged the median survival time of tumor-bearing animals compared with non-immunized animals (39, 33, 31 and 7 days, respectively). Both hemocyanins β-HlH and RvH activate the immune system of the experimental animals and therefore could be a good alternative for KLH. For this reason they could be included into the composition of non-specific anti-tumor vaccines to enhance their effectiveness.     

Relationship between classic histological pattern and sperm findings on fine needle aspiration map in infertile men

Systematic testis fine needle aspiration (FNA) mapping has been proposed as an adjunctive or alternative diagnostic procedure to biopsy to determine the presence of spermatozoa within infertile testes. This study related testis histology to the global presence or absence of spermatozoa in the same testes determined by FNA cytology. Testis biopsies and FNA mapping were performed in 87 infertile, azoospermic men. A mean of 1.3 biopsies and 14 FNA sites were taken per patient. Biopsies were assessed by recognized histological patterns of normal, Sertoli cell-only, hypospermatogenesis, early and late maturation arrest, sclerosis as well as mixed patterns that included at least two of these histologies. FNA cytological specimens were assessed for sperm presence by an experienced cytologist. Overall, spermatozoa were found by FNA mapping in 52% of patients. A comparison of histology and FNA findings revealed that pure patterns of Sertoli cell-only and early maturation were associated with a very poor likelihood of sperm detection (4-8%). In contrast, patients with other pure pattern histologies or mixed patterns had high rates of FNA sperm detection (77-100%). Similar to reported testicular sperm extraction (TESE) findings, sperm detection with FNA shows wide variation depending on testis histology. Unlike most TESE reports, however, some histological patterns generally reflect a more global testicular dysfunction and poorer likelihood of sperm identification, suggesting the possibility that these phenotypes have a genetic origin. Systematic testis sampling with FNA offers additional geographical information about spermatogenesis that routine biopsies lack and can further guide couple decision-making in severe male factor infertility.     

Lectin histochemistry of classic and spermatocytic seminoma

Six spermatocytic and 13 classic seminomas were studied histochemically with fluorescein isothiocyanate-labeled lectins. Wheat-germ lectin and succinyl concanavalin A reacted with all 19 tumors, whereas other lectins bound to some, but never to all, tumors in either group. Soybean lectin reacted with seven of 13 classic seminomas but with none of the spermatocytic seminomas. On the other hand, Maclura pomifera lectin reacted with two of six spermatocytic and with none of the classic seminomas. Our data thus illustrate some differences between classic and spermatocytic seminomas, but no diagnostic or pathognomonic pattern of lectin binding could be discerned. Neither classic nor spermatocytic seminomas (with a single exception) reacted with the lectins that do not react with normal spermatogenic cells. Spermatocytic seminomas did not react with eight lectins typically recognizing spermatids and spermatozoa. Thus, our data do not support the earlier contentions that spermatocytic seminomas contain malignant equivalents of cells in advanced stages of spermatogenesis.     

No significant association between HLA antigens and classic kaposi sarcoma: Molecular analysis of 49 jewish patients

Because of the abundance of the classical form of Kaposi sarcoma (CKS) among Jews and people of Mediterranean origin, studies have been conducted to find an association between CKS and HLA antigens. No conclusive results have been drawn, although in a number of these investigations an increased incidence of HLA-DR5 was reported. In our study 49 CKS patients of Jewish origin were serologically analyzed for HLA class I and class II antigens. We found no significant deviation in serologically defined HLA antigens frequencies between patients and 99 ethnically matched controls. However, a nonsignificant decrease in the frequencies of HLA-DR4, HLA-DR12, and the combination of DR4/DR11 was observed. Then we determined in these patients HLA-DRB1, HLADQA1, and DQB1 allelic polymorphism by oligotyping on PCR-amplified DNA. In this molecular analysis the only notable finding was a decreased frequency of the combination HLA-DQA1^*0301/DQB1^*0301, which appeared in one (2%) patient compared to 13 (13%) of the controls. However, the decrease was not statistically significant. On the basis of both serological and molecular analysis done on a relatively large group of CKS patients, we conclude that there is no significant linkage between HLA antigens and CKS in Jewish population.     

Clinical and phenotypic differences between classic and hypervirulent Klebsiella pneumonia: an emerging and under-recognized pathogenic variant

The purpose of this study was to increase awareness, gain insight into acquisition, and assess the virulence of the hypervirulent (hypermucoviscous) clinical variant (hvKP) that is entrenched in the Pacific Rim but emerging in Western countries. A case of community-acquired liver abscess with metastatic spread to the spleen is described. Comparative in vitro and in vivo virulence studies on this isolate (hvKP1) and four randomly chosen blood isolates of "classic" K. pneumonia strains (cKP1-4) were performed. Cases of hvKP infection are occurring in Western countries and are under-recognized. A hypermucoviscous phenotype is a surrogate laboratory marker for this variant. The propensity of hvKP strains for metastatic spread in non-compromised hosts is both a defining and unusual trait. The mode of acquisition in the described case was unclear but potential means are discussed. hvKP1 was more resistant to complement and neutrophil-mediated bactericidal activity and was more virulent in a rat subcutaneous abscess model than cKP1-4. Recognition of the hypermucoviscous phenotype, defined by a positive "string-test", will alert the microbiologist or clinician that the infecting strain may be a hvKP, which is hypervirulent compared to cKP. This will improve our understanding of the epidemiology and clinical spectrum of infection, which may be more extensive than appreciated.     

Relationship between classic Hodgkin lymphoma and overlapping large cell lymphoma investigated by comparative expressed sequence hybridization expression profiling

There is a diagnostic grey zone between classic Hodgkin lymphoma (cHL) and some non-Hodgkin lymphoma (NHL), including primary mediastinal B cell lymphoma, diffuse large B cell lymphoma, and anaplastic large cell lymphoma. They all have some morphological and/or phenotypic features in common. To investigate this, we undertook an expression profiling study of these lymphomas using comparative expressed sequence hybridization. This technique detects chromosomal regions that are differentially expressed between a test and a reference tissue in a manner similar to comparative genomic hybridization, and is particularly suitable when the number of informative biopsies is limited. Using this approach, we identified a unique expression profile for all lymphoma types investigated. Unsupervised hierarchical cluster analysis of the acquired data showed that cHL separates from all investigated NHLs, including ALCL-like HL. Moreover, anaplastic lymphoma kinase (ALK)-negative ALCL clustered in a separate branch together with ALCL-like HL. Thus, analysing the neoplastic cells concurrently with their microenvironment, ALK-negative ALCL and ALCL-like HL seem to be related to each other, while cHL constitutes a separate lymphoma entity.     

Transfusion-related acute lung injury investigation insights

Background From 2005 to 2009 transfusion-related acute lung injury (TRALI) has maintained its ranking as the number one cause of transfusion-related fatalities reported to the FDA. This confirms that TRALI remains a serious and potentially fatal transfusion complication. As over 80% of TRALI events have been attributed to donor derived leucocyte antibodies the detection and management of donors with these antibodies is crucial to reducing the TRALI risk. Objective Because blood is a precious medical commodity in limited supply, it would be more effective to exclude only those donors with a risk of triggering TRALI substantiated by objective laboratory evidence (i.e. implicated). This would allow implicated donors to be confidently excluded and would allow other donors only clinically associated with TRALI to be reassessed for continuing to donate. Therefore, the design of an effective and objective TRALI laboratory investigation strategy has to be based on current knowledge of the mechanism of antibody mediated TRALI. Discussion Leucocyte antibodies in the transfused blood product are thought to activate neutrophils in the pulmonary microvasculature. The by-products of neutrophil activation (e.g. reactive oxygen species and enzymes) consequently cause injury to the pulmonary microvasculature resulting in respiratory distress. There is strong evidence for the role of neutrophil reactive antibodies to human neutrophil antigen (HNA)-3a and human leucocyte antigen (HLA)-A2 in serious TRALI events. HLA Class II antibodies have also been implicated and they are thought to activate monocytes, which subsequently activate neutrophils. Neutrophils, which express human neutrophil antigens (HNA) and HLA Class I, are thus key effector cells in TRALI injury. Because of the pivotal role of neutrophils, effective TRALI investigations must include well validated neutrophil assays such as the granulocyte immunofluorescence test (GIFT) and granulocyte agglutination test (GAT). This article will discuss: – TRALI investigation strategy, – assays for detection of antibodies to HNA and HLA, – how to differentiate associated from implicated donors, – and provide thoughts on the remaining 20% of TRALI events (i.e. non-immune mediated).