IgA型自身抗体导致的自身免疫性溶血性贫血1例

目的通过使用微柱凝胶卡式法进行Coombs试验,确定导致自身免疫性溶血性贫血患者的自身抗体类型,从而在临床输血过程中注意此种情况,减少输血反应。方法用微柱凝胶卡式法检测患者直接抗人球蛋白试验,分别用微柱凝胶检测卡法和凝聚胺法对患者进行主、次侧交叉配血试验,以明确该类患者在交叉配血时所需采用的试验方法。结果患者DAT试验结果为阳性,具体抗体分型为IgA型;微柱凝胶法交叉配血患者主、次侧交叉配血试验结果均为阴性;凝聚胺法主、次侧交叉配血结果均为阳性。结论单独型IgA抗体的AIHA患者,除微柱凝胶检测卡交叉配血外,还需用凝聚胺法进行交叉配血来共同参考配血结果。     

微柱凝胶技术在新生儿溶血病抗体检测中的应用价值

目的 探讨微柱凝胶法在新生儿溶血病抗体检测中的应用价值,为临床诊断提供依据.方法 对249例新生儿溶血病患儿,分别应用微柱凝胶法、试管法进行直接抗人球蛋白试验、抗体游离试验及抗体放散试验检测,并对检测结果进行对比分析.结果 微柱凝胶法检出直接抗人球蛋白试验阳性率为90.4％,抗体游离试验阳性率为92.8％,抗体放散试验阳性率为96.4％;试管法检出直接抗人球蛋白试验阳性率为73.5％,抗体游离试验阳性率为80.7％,抗体放散试验阳性率为85.9％;微柱凝胶法检出阳性率均显著高于试管法(P＜0.01).结论 微柱凝胶法对检测新生儿溶血病抗体的检出率高于试管法,值得临床推广应用.     

自身免疫性溶血性贫血的微柱凝集技术诊断应用

目的探讨微柱凝集体技术在诊断自身免疫性溶血性贫血(AIHA)中的应用.方法采用BioVue系统微柱凝集技术对26例AIHA患者进行检测,同时用传统的试管法抗人球蛋白试验作比较.结果统计学上两种方法对AIHA检出率无显著性差别,但微柱凝集技术能检出试管法所不能检出的阴性血样,比试管法检测凝集强度高1+～2+.结论BiVue系统微柱凝集技术能快速准确检测出AIHA,具有更高的灵敏度,是直观可靠的方法.     

微柱凝胶技术在自身免疫性溶血性贫血实验诊断中的应用

目的应用微柱凝胶检测技术探讨自身免疫性溶血性贫血的实验诊断依据。方法采用试管法直接抗人球蛋白试验、微柱凝胶抗人球蛋白试验对住院怀疑为免疫性溶血的患者进行检测。结果受检的102份血样中,试管法直接抗球蛋白试验阳性14例,阳性率凝胶直接抗球蛋白试验阳性19例。两种检测方法间比较差异有统计学意义（χ2=7.14,P〈0.05）。结论微柱凝胶抗人球蛋白试验是一种免疫学检测的新技术,对自身免疫性溶血性贫血患儿的诊断,微柱凝胶抗人球蛋白试验比试管法具有更多优点。微柱凝胶技术具有操作简便、结果易判读,敏感性高,特异性强,结果可靠等优点。     

凝胶微柱法检测酸放散液中的抗体

试管法作为红细胞(RBC)血清学试验中检测抗体的标准方法已有数十年,然而随着凝胶微柱抗体检测系统的大量运用,许多试验可以用凝胶微柱法代替试管法。凝胶微柱法已用于检查ABO、Rh血型、血清中的意外抗体、进行直接抗人球蛋白试验(DAT)。与试管法相比,凝胶微柱法只需少量的样本和试剂,结果判读简便且更易于自动化,因此许多血液中心及输血机构都采用此法。本试验的目的在于比较凝胶微柱法和试管法检测红细胞放散液中抗体的能力,放散液则来源于脐带血和外周血经上述两种方法检测DAT阳性或DAT阴性的红细胞。     

凝胶微柱法与试管法检测放散液中抗体的比较

目的比较凝胶微柱法和试管法检测红细胞释放液抗体的能力。方法将30例外周血和41例脐血红细胞用两种方法测定直接抗人球蛋白试验(DAT)并进行酸放散,将放散液与三种谱细胞、A1、B细胞反应检测抗体的特异性。71例中12例作为阴性对照样本(DAT均为阴性),其中6例为健康献血者外周血,6例为不发生新生儿溶血病(Heamolytic disease of thenewborn,HDN)的婴儿脐血样本。结果24例外周血放散液中有10例两种方法均阳性,12例均为阴性,2例自身免疫性溶血性贫血(AIHA)患者样本凝胶微柱法阳性而试管凝集法阴性,6例健康捐献者的样本DAT试验阴性且两种方法检测放散液均为阴性;41例脐血样本中33例放散液两种方法均反应,2例放散液在试管法中与A1和B细胞反应而同样的放散液做凝胶微柱法时一个和A1细胞反应,另一个只与B细胞反应。结论两种方法检测放散液结果基本是一致的,检测AIHA患者红细胞释放液抗体时用微柱法比较好,检测脐血细胞释放的同种血凝素时试管凝集法更好—些。     

ABC-ELISA检测AIHA患者EAIgG的结果分析

目的探讨亲和生物素化酶复合物-酶联免疫吸附试验(ABC-ELISA)法定量检测红细胞结合IgG(EAIgG)在自身免疫性溶血性贫血(AIHA)中的诊断价值.方法采用ABC-ELISA法定量检测EAIgG.共检测36名正常对照、38例AIHA患者(其中6例Coombs′试验阴性的AIHA患者)的EAIgG值,并同时与直接抗人球蛋白试验(DAT)结果比较.结果 AIHA患者的EAIgG值显著增高,与正常对照组相比,差异有显著性(P<0.001);6例Coombs′试验阴性的AIHA患者EAIgG均值(3.93±1.77)fg/RBC,Coombs′试验阳性的AIHA患者EAIgG均值(2.89±1.72)fg/RBC,两者相比差异无显著性(P>0.05).结论 ABC-ELISA法定量检测EAIgG有助于AIHA的诊断,特别对Coombs′试验阴性的AIHA患者更具确诊价值.     

ABC-ELISA检测AIHA患者EAIgG的结果分析

目的　探讨亲和生物素化酶复合物 酶联免疫吸附试验 (ABC ELISA)法定量检测红细胞结合IgG(EAIgG)在自身免疫性溶血性贫血 (AIHA)中的诊断价值。 方法　采用ABC ELISA法定量检测EAIgG。共检测 36名正常对照、38例AIHA患者 (其中 6例Coombs′试验阴性的AIHA患者 )的EAIgG值 ,并同时与直接抗人球蛋白试验 (DAT)结果比较。结果　AIHA患者的EAIgG值显著增高 ,与正常对照组相比 ,差异有显著性 (P <0 .0 0 1) ;6例Coombs′试验阴性的AIHA患者EAIgG均值 (3.93± 1.77)fg/RBC ,Coombs′试验阳性的AIHA患者EAIgG均值 (2 .89± 1.72 )fg/RBC ,两者相比差异无显著性 (P >0 .0 5 )。 结论　ABC ELISA法定量检测EAIgG有助于AIHA的诊断 ,特别对Coombs′试验阴性的AIHA患者更具确诊价值。     

自身免疫性溶血性贫血的微柱凝集技术诊断应用

目的　探讨微柱凝集体技术在诊断自身免疫性溶血性贫血 (AIHA)中的应用。方法　采用BioVue系统微柱凝集技术对 2 6例AIHA患者进行检测 ,同时用传统的试管法抗人球蛋白试验作比较。结果　统计学上两种方法对AIHA检出率无显著性差别 ,但微柱凝集技术能检出试管法所不能检出的阴性血样 ,比试管法检测凝集强度高 1 ～ 2 。结论　BioVue系统微柱凝集技术能快速准确检测出AIHA ,具有更高的灵敏度 ,是直观可靠的方法。     

ABO血型不合新生儿溶血病3种检测方法的比较

目的比较ABO血型不合新生儿溶血病(HDN)传统试管检测法、凝胶卡式检测法和流式细胞仪检测法的优缺点。方法对临床拟诊的110例HDN标本同时用试管法、凝胶卡式法和流式细胞仪对患儿红细胞直接抗球蛋白试验(DAT)、血清游离抗体检测及患儿红细胞放散液鉴定3项试验进行检测,并对检测结果进行比较。结果110例HDN标本用试管法检测出符合ABO-HDN血型血清学特征的阳性标本55例,阳性率50%;凝胶法检测阳性73例,阳性率66%;流式细胞仪检测阳性95例,阳性率86%。结论3项试验的3种方法检测灵敏度从高到低排列依次是流式细胞仪法、凝胶卡式法、试管法;流式法的测定数值客观、定量、准确,有利于结果的判断,凝胶法操作简单快速,结果稳定且易保存,2种方法相对于传统的试管法具有明显的优点。     

Comparison of conventional tube test technique and gel microcolumn assay for direct antiglobulin test: a large study

Gel microcolumn assay (GMA) is a modified serological technique that has been used for ABO and Rh typing, direct antiglobulin test (DAT), detecting alloantibodies, red cell phenotyping, and other applications. However, for DAT, the role of GMA is controversial. The purpose of this large study was to compare the performance of the conventional tube test (CTT) to GMA for detecting potentially significant antibodies coating red blood cells in vivo. From January 1996 to May 2002, we performed DATs by GMA and CTT on 9,862 blood samples submitted to our reference laboratory, using LISS/Coombs cards (DiaMed-Latino America, Lagoa Santa-MG, Brazil) for GMA and polyspecific and monospecific anti-IgG reagents for CTT. Acid eluates were prepared from all positive DAT samples. The specificity of eluates was determined by GMA. We detected nonconcordant results in 2,079 out of 3,163 positive DATs (65.7%). All of these tests were only positive in GMA. Sensitivity and specificity for DATs was 100% and 83.0% for gel, and 50.7% and 97.8% for tube, respectively. Based on this study GMA showed to be more sensitive than CTT for detecting potentially significant antibodies coating red blood cells in vivo.     

ABC—ELISA法定量检测红细胞相关IgG及临床意义

目的 寻求一种温抗体型自身免疫性溶血性贫血（ＷＡＩＨＡ）的诊断方法,以期提高ＷＡＩＨＡ的诊断率。方法 采用本室建立的亲和素生物素化酶复合物酶联免疫吸附测定（ＡＢＣ－ＥＬＩＳＡ）,定量检测红细胞相关ＩｇＧ（ＥＡＩｇＧ）。共中正常人、１９６例ＷＡＩＨＡ及各类疾病患的ＥＡＩｇＧ值,并与Ｃｏｏｍｂｓ试验作比较。结果 ＷＡＩＨＡ组患的ＥＡＩｇＧ显增高,与正常组、非免疫性贫血组及免疫性疾病组的ＥＡＩｇＧ     

Use of gel microcolumn assay for the detection of drug-induced positive direct antiglobulin tests

Drugs can result in broad variety of hematologic abnormalities including positive direct antiglobulin test. In this study, we evaluated gel microcolumn assay for the detection of drug-induced antibodies. Direct antiglobulin test was performed by conventional tube and by gel microcolumn assay in 139 hospitalized patients. Drug in vitro studies were done in 34 patients with positive direct antiglobulin test by tube test and gel microcolumn assay using serum and eluate. None of them had signs of hemolytic anemia. A total of 1,000 blood samples from donors were used as control group. Gel microcolumn assay was more sensitive than in tube test for direct antiglobulin test (P<0.01). Positive direct antiglobulin test was more frequent in patients than in donors (P<0.01). Drug in vitro studies were positive with at least one drug in 76.5% of patients with positive direct antiglobulin test by immune complex and/or adsorption mechanisms. We found a high incidence of positive drug in vitro tests in positive direct antiglobulin test patients. Gel microcolumn assay showed appropriate results for drug in vitro studies. The combination of tube and gel microcolumn assay can improve detection of drug-induced positive direct antiglobulin tests.     

首发症状Coombs试验阴性自身免疫性溶血性贫血的非何杰金淋巴瘤(英文)

非何杰金淋巴瘤(NHL)伴发自身免疫性溶血性贫血(AIHA)或AIHA发生在NHL诊断之前或治疗过程中已有不少报道,但以Coombs试验阴性AIHA为首发症状的NHL鲜有报道。在此,本文报道1例1.5年反复溶血发作的Coombs试验阴性AIHA后合并NHL的患者。患者女性,69岁,根据病史和实验室检查诊断为Coombs试验阴性AIHA,给予强的松治疗后血红蛋白恢复正常,停药后溶血反复复发2次,强的松治疗仍有效。第3次复发时强的松治疗无效,并出现颈部淋巴结肿大,病理检查确诊为NHL。给予6个疗程的CHOP方案化疗,NHL治愈,但溶血仍不能控制,给予小剂量利妥昔单克隆抗体(rituximab,RTX)治疗后,溶血很快停止,此后给予3次小剂量RTX进行维持治疗,NHL和AIHA呈持续缓解状态。结论:本文报告了一例十分罕见的非何杰金淋巴瘤,其发病时的主要临床症状为Coombs阴性自身免疫性溶血性贫血。     

Diagnosis and treatment of autoimmune haemolytic anaemias in adults: a clinical review

Autoimmune haemolytic anaemia (AIHA) is an immune disorder caused by antibodies directed against unmodified autologous red cells. The disorder may be a primary (idiopathic) or a secondary disease. The diagnosis is based on the presence of anaemia, signs of haemolysis with reticulocytosis, low haptoglobin, increased lactate dehydrogenase, elevated indirect bilirubin, and a positive direct antiglobulin test (Coombs test). Sometimes, not all of these typical features are present. Most AIHA are caused by warm antibodies, whereas cold antibodies are less commonly detected. While half of the warm antibody-based AIHA are idiopathic anaemias, almost all cold antibody AIHA are secondary anaemias. Underlying diseases are Non Hodgkin's lymphomas and systemic autoimmune disorders, and less frequently organ transplantation, infections, or solid tumors. Moreover, AIHA is an important complication of treatment with nucleoside analogs. Most patients with AIHA require therapy. In warm antibody AIHA, standard first line therapy are glucocorticosteroids with or without high dose immunoglobulins, whereas splenectomy is considered second-line therapy. Response rates of primary AIHA to corticosteroid therapy are high. After initial remission, the dose should be tapered down slowly and with caution, and in some cases, low-dose maintenance therapy is required. The efficacy of standard therapy is low in secondary AIHA that develops in lymphoma patients, posttransplant patients, or tumor patients. Among other immunosuppressive treatments, rituximab (anti-CD20) appears to be highly effective in patients with warm antibody AIHA refractory to standard therapy. Mycophenolate mofetil is quite effective in AIHA patients with an underlying autoimmune or lymphoproliferative disease. Patients with cold agglutinins are refractory to steroids and splenectomy. Half of these patients may respond to rituximab, although responses usually are short-lived. Sometimes, AIHA that is associated with malignant lymphomas or tumors, disappears after successful anti-lymphoma or anti-tumor therapy.     

DNA testing for sickle cell anemia in Africa: Implementation choices for the Democratic Republic of Congo

BackgroundHemoglobin‐based tests form the reference diagnostic test for SCA. In limited resource countries, these tests face limitations including cost, low sensitivity due to recurrent transfusions in endemic malaria region, and interference from fetal hemoglobin in neonatal diagnostic. This study aimed at adapting DNA‐based SCA tests to limited resource countries and evaluating the economic benefit.Methods338 participants were recruited in the Democratic Republic of Congo, sorted in 3 cohorts based on venous blood, umbilical cord blood (UCB) and buccal swab sampling. RFLP was performed to identify mutated allele. The feasibility and technical validity of this RFLP was evaluated for specimens collected on DBS cards and on EDTA tubes. RFLP on DBS stored at room temperature was regularly repeated to assess sample conservation. Finally, the cost analysis was performed.ResultsDBS cards yielded identical results to extracted DNA. Repeated testing returned the same result after four years. The DBS‐based test performed on UCB or on buccal swab had a sensitivity and a precision of 100%. Cost comparison indicated that our approach costs half price of the widely used isoelectrofocussing of hemoglobin.ConclusionThe implemented DNA‐based test approach overcomes the limitations faced by hemoglobin‐based tests, while being more affordable. We propose to implement the RFLP test as a first line diagnostic test after transfusion and as second tiers for newborn screening. However, users should be aware that this test is unable to differentiate HbC from HbS or identify other point mutation of gene deletion of HBB gene.     

False Positive Coombs Tests Due to Bacterial Contaminants in Stored Pilot Tube Blood

Unexplained positive Coombs tests during crossmatches have occasionally occurred in this laboratory. A brief investigation was carried out to determine if bacterial contamination of pilot tube blood was involved in this phenomenon. Of 220 pilot tubes cultured, seven contained viable bacteria. These organisms were inoculated into sterile bloods and incubated. Coombs tests were then performed. Blood containing Pseudomonas effusa gave positive direct and indirect antihuman globulin reactions, and blood containing Bacillus subtilis gave positive indirect antihuman globulin reactions. Blood containing other organisms isolated in this study gave negative results with Coombs tests.     

Clinical application of a flow cytometric direct antiglobulin test

BACKGROUND: The clinical application of flow cytometric direct antiglobulin test (FC-DAT) has rarely been evaluated for patients with various diseases including immune and nonimmune hemolytic anemia.
STUDY DESIGN AND METHODS: Blood samples from 380 patients with a variety of diseases were studied using the tube direct DAT and FC-DAT. The results of tube DAT and FC-DAT were compared. The predictive values of DAT for hemolysis were evaluated.
RESULTS: Of 57 patients with autoimmune hemolytic anemia (AIHA), 6 of the 17 with a negative tube DAT (immunoglobulin G [IgG]) had a positive FC-DAT (IgG) and 23 of the 36 patients with a negative tube DAT (complement 3d [C3d]) had a positive FC-DAT (C3d). In 57 patients with AIHA, the incidence of positive results of FC-DAT (IgG) and tube DAT (IgG) were similar (42 positive vs. 40 positive); but in 323 patients without AIHA, the incidence of positive FC-DATs (IgG) was higher than that of tube DAT (IgG; 47 positive vs. 9 positive). The higher incidence of positive FC-DAT (C3d) than that of tube DAT (C3d) was seen in patients with AIHA (42 positive vs. 21 positive) as well as in patients without AIHA (61 positive vs. 5 positive). Both DAT (IgG) and DAT (C3d) positive has highest positive predictive value for hemolysis, followed by DAT (IgG) alone positive and DAT (C3d) alone positive.
CONCLUSIONS: FC-DAT is a complementary test for diagnosing AIHA. There is a synergistic effect of the red blood cell–bound IgG and complement in predicting hemolysis.     

伴有自身免疫性溶血性贫血及纯红系再生障碍性贫血的血管免疫母细胞性T细胞淋巴瘤

血管免疫母细胞性T细胞淋巴瘤（AITL）是一种外周T细胞淋巴瘤,常合并自身免疫现象,如免疫相关性血细胞减少症,是NHL中的少见类型。为了研究AITL的临床特征,病理表现和有效的治疗方法,对1例37岁男性患者进行了血常规检查、骨髓检测、单个核细胞的流式细胞术检测、Coombs试验、血清学检测、CT和免疫组织化学测定等。结果查明,患者有广泛淋巴结肿大、肝脾肿大,颈部淋巴结活检表明为血管免疫母细胞性T细胞淋巴瘤;患者重度贫血,网织红细胞降低,Coombs实验阳性,骨髓红系增生低下,提示并发温抗体型自身免疫性溶血性贫血（AIHA）和纯红系再生障碍性贫血（PRCA）;经过CHOP-E方案化疗后,合并的AIHA和PRCA以及AITL浸润症状均消失。结论：成功地确诊了合并有AIHA和PRCA的AITL,淋巴结活检和骨髓检测意义大,CHOP-E化疗方案对此种AITL有一定治疗效果。     

微柱凝胶法在新生儿溶血病检测中的应用

目的采用微柱凝胶技术进行新生儿溶血病血清学检测,对新生儿直接抗人球蛋白试验、游离试验、放散试验的结果进行分析。方法采用微柱凝胶技术对新生儿溶血病进行检测。结果在108例送检的标本中74例确诊为新生儿溶血病,占送检标本的68．5％（74／108）;出生天数小于或等于5d者新生儿溶血病检出率为72．3%（68／94）,明显高于出生天数大于5d的新生儿42．9％（6／14）。结论微柱凝胶技术与传统试管法相比,具有操作简便、敏感性高、准确等优点;新生儿外周血标本的早期检测,可提高阳性指标的检出率。