抗体放散试验在直接抗人球蛋白试验阳性患者中的应用

目的探讨抗体放散试验在直接抗人球蛋白试验(DAT)阳性患者输血前实验室检查中的意义,为临床输血安全提供保障。方法通过回顾性分析74例患者的DAT阳性强弱度、放散液和血清中抗体特异性比较、经稀释后的放散液中的抗体特异性,探讨放散试验在输血前检查中的应用。结果 74例DAT阳性反应强弱度为:W+(13.5%),1+(31.1%),2+(32.4%),3+(16.2%),4+(6.8%);放散后抗体鉴定32例为阴性,16例放散液检测到特异性同种抗体,和患者血清中的一致,并且所有患者均有输血史;26例放散液为全凝集,进行了稀释,其中4例检测到了同种抗体。结论抗体放散试验在输血前检查中并为常规试验,但对于DAT阳性的患者来说进行放散试验是很有必要的。     

自身抗体阳性伴直接抗人球蛋白试验阳性患者血型血清学特征及输血疗效

目的探讨自身抗体和直接抗人蛋白试验(DAT)阳性患者的血型血清学特征及配血的处理方法,寻求安全、有效的输血对策。方法结合吸收放散方法采用不同厂家、批号的抗筛细胞和谱细胞对待检血清和红细胞放散液进行自身抗体和同种抗体特异性的鉴别;选取合适的献血者进行主次侧交叉配血试验;分析自身抗体、同种抗体的特异性及其与ABO血型、疾病、贫血的关联和对输血及输血疗效的影响。结果 139例研究对象均为自身抗体阳性伴DAT阳性且交叉配血试验主次侧均不合,其中20例只含有自身抗体,59例伴Rh系统或MNs系统或Kidd系统抗体,47例为自身抗体不能排除同种抗体,13例伴有药物抗体或其他系统抗体;共进行221次输血治疗,未发生溶血性输血反应。自身抗体和DAT的凝集强度与贫血呈正相关(P0.05),与输血疗效呈负相关(P0.05);自身抗体的强度与自身红细胞被致敏的程度呈正相关(P0.05),且多为免疫性疾病,与ABO血型无明显相关性(P0.05)。结论对于自身抗体伴DAT阳性的病例,其贫血的严重程度、输血次数、输血后疗效与自身抗体和DAT的凝集强度有关,交叉配血前必须采用合理的流程和方法判定是否伴有同种抗体,选取相应抗原阴性的献血者或该地区出现较高频率的同种抗体对应的抗原系统同型的献血者配血,可提高安全输血效率,最大限度地减少输血不良反应发生。     

输血前不规则抗体筛查与直接抗人球蛋白试验的意义

目的分析永州地区住院输血前患者不规则抗体及直接抗人球蛋白试验(DAT)的阳性率及抗体特异性。方法选择3454例住院输血前患者,采用微柱凝胶法筛查不规则抗体及DAT,并对筛查阳性标本进行抗体特异性鉴定。结果在3454例患者中共检测出不规则抗体6例,阳性率为0.17%,其中抗-E3例、抗-C1例、抗-c1例、抗-JKa1例,DAT阳性35例,阳性率为1.01%,其中IgG型抗体18例、C3型抗体7例、IgG+C3型抗体10例。结论输血前筛查不规则抗体及DAT,在保证临床输血安全、减少溶血性输血反应方面具有重要的意义。     

不规则抗体阴性受血者直接抗人球蛋白试验阳性的特征分析及类型鉴别

目的 不规则抗体阴性受血者直接抗人球蛋白试验阳性的特征分析及类型鉴别.方法 将2019年9月至2020年4月在该院行不规则抗体筛查,结果为阴性的受血者(2066例)中交叉配血次侧凝集的受血者72例纳入研究作进一步分析.对交叉配血次侧凝集的受血者行DAT、分型试验、热放散试验及输血疗效评估,分析其分布特征.结果 72例不规则抗体筛查阴性交叉配血次侧凝集的受血者均为DAT阳性;Hb>50～60 g/L的受血者所占比例最高(P<0.05),为40.28％;输血次数≥3次,所占比例最高(P<0.05),为37.50％;输注有效占79.17％,输注部分有效的占15.28％,输注无效的占5.56％;DAT阳性类型的分布:单纯IgG型占91.67％,IgG+C3d型占2.78％,分型未果占5.56％.将上述72例受血者的DAT阳性标本行热放散试验后再进行抗体筛查,结果为全阴性的占79.17％(57/72),全阳性的占15.28％(11/72),抗体筛查有反应格局的占5.56％.结论 DAT阳性可导致输血无效,应重视和加强此类受血者DAT阳性类型的鉴别,并结合患者的临床情况进行全面分析,预防输血不良反应的发生.     

微柱凝胶法检测头孢唑啉钠和头孢西丁钠诱导形成药物性抗体的可行性研究

目的研究微柱凝胶法检测头孢唑啉钠和头孢西丁钠诱导形成药物性抗体的可行性。方法采用微柱凝胶法检测经头孢唑啉钠和头孢西丁钠治疗168例患者的血液标本,进行间接抗人球蛋白试验(IAT)与直接抗人球蛋白试验(DAT),并检测对应药物性抗体。结果 IAT结果显示,168例血浆标本中不规则抗体筛查结果均未出现阳性;DAT结果显示,168例血浆标本中DAT结果阳性32例(19.05%),其中采用头孢唑啉钠治疗的患者阳性20例,采用头孢西丁钠治疗的患者阳性12例;头孢唑啉钠药物性抗体阳性率高于头孢西丁钠(P0.05);头孢唑啉钠DAT结果阳性标本与头孢西丁钠DAT结果阳性标本中药物性抗体阳性率比较,差异无统计学意义(P0.05);头孢唑啉钠DAT结果阴性标本中药物性抗体阳性率高于头孢西丁钠DAT结果阴性标本(P0.05)。头孢唑啉钠、头孢西丁钠DAT结果阳性标本中不规则抗体检测结果均为阴性,药物性抗体均为阳性。结论微柱凝胶法检测头孢唑啉钠和头孢西丁钠诱导形成药物性抗体时操作简便,结果稳定,可行性强,可用于临床检验工作。     

临床患者血浆中β- 内酰胺类药物＆抗体致输血无效及对供者红细胞亲和力的研究

目的 分析临床患者血浆中β- 内酰胺类药物＆抗体对供者红细胞的亲和力,探究药源性溶血性贫血及输血无效的临床特点。方法 选择2021 年11 月～ 2022 年4 月期间临床送检的4 例有β- 内酰胺类药物用药史的临床患者,检测ABO 与Rh 血型,直接抗球蛋白试验（direct antiglobulin test ,DAT）、不规则抗体鉴定（identification of irregularantibodies,IAT）与β- 内酰胺类药物抗体及效价,对DAT 阳性的患者红细胞进行酸放散并检测放散液中β- 内酰胺类药物抗体。选择ABO 和Rh 同型的供者红细胞与患者血浆在37℃无菌条件下体外致敏,监测β- 内酰胺类药物＆抗体在0 ,24,48 和72 h 与供者红细胞的亲和力,并观察加入补体后的溶血程度。结果 4 例患者血浆中均存在β- 内酰胺类药物抗体,停药前输血无效,停药后输血效果良好。患者1:A 型,RhCCDee,DAT 阳性（3+W）,IAT 阴性,血浆中存在头孢哌酮药物抗体（效价:1∶64）、阿莫西林药物抗体（效价:1∶16）,放散液中存在头孢哌酮药物抗体。患者2:A 型,RhCcDee,DAT 阳性（4+W）,IAT 阴性,血浆中存在头孢哌酮药物抗体（效价:1∶128）,放散液中存在头孢哌酮药物抗体。患者3:A 型,RhCcDee,DAT 阳性（4+）,IAT 阳性,鉴定为抗 E 抗体,血浆中存在阿莫西林药物抗体（效价:1∶16）、亚胺培南药物抗体（效价:1∶64）、头孢哌酮药物抗体（效价:1∶128）,放散液中存在亚胺培南、头孢哌酮药物抗体。患者4:A 型,RhCCDee ,DAT 阳性（1+）,IAT 阳性,鉴定为抗-E.c 抗体,血浆中存在阿莫西林药物抗体（效价:1∶32）,放散液中未检测到药物抗体。4 例患者血浆与供者红细胞体外致敏24 h 后DAT均为阳性,48 和72 h 内逐渐增强,加入补体后均可引起红细胞溶解。结论 头孢哌酮、阿莫西林和亚胺培南三种β-内酰胺类药物＆抗体可迅速结合到供者红细胞表面并随时间延长而增强,有补体存在的条件下可在24 ～ 72 h 内破坏供者红细胞引起溶血,导致输血无效。     

新生儿溶血病检测试验多样性与胆红素水平相关性研究

目的探讨新生儿ABO溶血病（ABO．HDN）溶血三项试验不同结果与胆红素水平的相关性。方法对85例ABO—HDN患儿进行血清溶血三项试验[抗人球蛋白试验（DAT）、游离抗体试验、抗体释放试验]、母亲IgG抗A（B）抗体、血清胆红素等测定。结果ABO．HDN患儿DAT阳性者血清总胆红素峰值与DAT阴性者比较,差异有统计学意义（P〈0．05,0．01）;DAT阴性间比较、DAT阳性问比较（P〉0．05）,差异无统计学意义。新生儿ABO．HDN患儿母亲血型抗体与血清总胆红素水平比较,差异无统计学意义（P〉0．05）。结论ABO-HDN患儿DAT阳性合并抗体释放试验阳性或三项试验均阳性时,血清胆红素水平增高较明显,DAT阴性释放试验和／或游离试验阳性时,新生儿ABO-HDN相对较轻,而母亲IgG抗体效价高低不能反映HDN的严重程度,这可为临床合理治疗HDN提供帮助。     

不规则抗体筛查阳性结果分析及输血策略

目的通过对输血前不规则抗体筛查阳性结果分析,探讨安全有效的输血治疗策略。方法选择该院有输血史、妊娠史而需输血的1 896例住院患者为研究对象,采用微柱凝胶法,对所有患者进行不规则抗体筛查和鉴定。结果拟输血的1 896例患者中不规则抗体筛查阳性22例,阳性率为1.16%(22/1 896)。特异性同种抗体阳性17例,占77.27%(17/22),阳性率为0.90%(17/1 896)。其中Rh血型系统抗体15例,以抗-E检出率最高;MNS血型系统抗体中抗-M共1例;Lewis血型系统抗体中抗-Lea1例。抗体筛查阳性患者均选择对应抗原阴性的红细胞进行输注,所有输血患者取得了较好的输血效果,临床上均无输血不良反应发生。非特异性自身抗体阳性5例,占22.72%(5/22),经药物治疗后,患者贫血症状得到有效改善。结论通过对不规则抗体阳性结果的分析,选择合适的血液输注,可同时对于能用药物改善贫血症状的患者,可以用药物部分替代输血治疗,能有效地避免或降低免疫溶血性输血反应的发生,保证临床用血的安全。     

非自身免疫性溶血性贫血患者直接抗人球蛋白试验阳性对临床输血效果影响

目的:研究非AIHA患者DAT阳性对临床输血效果影响.方法:收集多个科室96例DAT阳性的非AIHA输血者为阳性组,均分为甲组(48例输注洗涤红细胞)和乙组(48例输注普通红细胞悬液),对照组为58例DAT阴性的常规输血患者.采用微柱凝胶法(MGT)和凝聚胺法进行交叉配血.检测患者红细胞(RBC)、血红蛋白(Hb)、红...     

直接抗人球蛋白试验阳性患者的疾病分布及输血疗效评价

目的 探讨直接抗人球蛋白试验（DAT）阳性患者的免疫分型与临床疾病分布特点,并评价其输血疗效。方法 回顾性分析1 204例于我院输血科进行DAT检测患者的临床资料,选择交叉配血过程中次侧配血不合的244例DAT阳性患者（未包含新生儿溶血病）为试验组,随机选择同时间段内配血相合、DAT阴性的100例患者（未包含新生儿溶血病）为对照组。试验组采用微柱凝胶技术进行DAT检测和免疫分型;比较各组红细胞输注疗效。结果 244例DAT阳性患者中,DAT凝集强度W+～2+占比83.20%,3+～4+占比16.80%。244例DAT阳性患者中,自身免疫性疾病占比最高,其次是血液系统疾病和肿瘤,其他疾病类型占比较低;肿瘤、感染性疾病和不明原因贫血患者DAT免疫分型主要以单纯抗体免疫球蛋白G(IgG)为主,单纯补体C3d次之;自身免疫性疾病和血液系统疾病的患者以单纯抗体IgG和IgG+C3d复合阳性较多。IgG组、C3d组输血后红细胞（RBC）、血红蛋白（Hb）、血细胞比容（Hct）变化值分别与对照组比较,差异无统计学意义（P>0.05）,而IgG+C3d组输血后RBC、Hb、Hct变化值明显小于对...     

Retrospective analysis of direct antiglobulin test positivity at tertiary academic hospital over 10 years

BackgroundHemolytic disease of the fetus and newborn (HDFN) is a clinically significant problem that may potentially affect any pregnancy. Direct antiglobulin test (DAT) is considered to be an important test in identifying newborns who are suspected to have HDN. This study aims in reviewing data regarding a positive DAT result concerning etiology and the development of HDN over a period of 10 years.Study design and methodsA retrospective study of all neonates with a positive DAT result between January 2011 and December 2020 was performed. Data were obtained from patients' electronic hospital files, transfusion medicine databases, and medical birth records. Laboratory parameters along with clinical interventions in neonates with a DAT-positive result and a comparison group of DAT-negative neonates were performed.Results36,000 deliveries were registered in this period. 176 (2.65 %) neonates had a positive DAT result. ABO-incompatibility was the most common cause with 59.1 %; Rh incompatibility 13.8 %, minor blood group incompatibility, and other RBC-related antibodies 10.1 %, and unspecified etiology in 17 % of cases. Among DAT-positive cases, 32.7 % of neonates were diagnosed with HDN. ABO-incompatibility was the major reason as well. Initial mean total bilirubin levels were higher in the DAT-positive group than the control group (p < 0.001), and these neonates also had a lower initial hemoglobin level (p < 0.001). The need for therapeutic interventions was significantly higher in DAT-positive neonates (p < 0.001) as 86.8 % underwent phototherapy, with 32.7 %, and 17.6 % receiving exchange transfusion (ET) and intravenous immunoglobulin (IVIG), respectively.ConclusionIn conclusion, ABO incompatibility was the most common cause for neonatal DAT positivity. Besides the common causes of DAT positivity, there would be rare but important conditions that may lead to a positive result, such as antibodies passively acquired from mothers in the context of alloimmunizations or using drugs. In addition, as a high rate of therapeutic intervention was identified among neonates with a DAT-positive result, there is a crucial need for increasing awareness regarding early diagnosis of the condition, careful monitoring, and the employment of prenatal alloimmunization screening tests.     

Lower hemoglobin levels in human immunodeficiency virus-infected patients with a positive direct antiglobulin test (DAT): relationship with DAT strength and clinical stages

BACKGROUND: There are conflicting opinions regarding the effect of positive direct antiglobulin test (DAT) on hemoglobin (Hb) levels in human immunodeficiency virus-infected (HIV+) patients. STUDY DESIGN AND METHODS: A total of 166 samples from HIV+ outpatients were studied. The DAT was performed with the tube test and column agglutination technology (CAT). RESULTS: The DAT was positive in 18.67 percent with the tube method and 33.73 percent with the CAT. Patients with DAT-positive results showed lower Hb levels than DAT-negative patients, 12.3 g per dL versus 14.3 g per dL (p = 0.0002). The univariate logistic regression enabled us to study the phenomenon better and fit the probability of having a DAT-positive result on the basis of the Hb levels. The relationship between the CAT and the tube test when washing the red blood cells (RBC) at 4 degrees C was stronger than when washing these at room temperature (phi = 0.8156; p = 0.000). The Hb levels were significantly lower in the positive DATs of Stage C (acquired immune deficiency syndrome [AIDS]) and Stage B (symptomatic non-AIDS patients), which showed decreasing Hb values for increasing agglutination strengths (p = 0.000). Anemia was related with the DAT results (odds ratio [OR], 8.005; p = 0.000) but not to the AIDS condition (OR, 1.741; p = 0.221). DISCUSSION: Our study indicates that the DAT-positive results may be specifically related to lower Hb levels in HIV+ patients. The immunologic RBC clearance could be part of the anemic multifactorial condition in HIV+ patients.     

输注辐照血小板后血液病患者免疫参数变化的临床观察

目的研究输注辐照血小板后患者体液免疫参数的变化以及这些免疫参数与血小板无效输注（PTR）的相关性。方法选取多次输血的恶性血液病患者随机分为观察组和对照组,分别输注γ辐照机采血小板和普通机采血小板,比较两组输注的有效率和输血不良反应的发生率,检测输血前后免疫球蛋白（IgG、IgM、IgA）、补体（C3、C4）、循环免疫复合体（CIC）、C反应蛋白的变化。分析发生无效输注患者的免疫参数特点。结果输注机采血小板后血清免疫球蛋白（IgG、IgM）、补体（C3、C4）、CIC在观察组变化差异不大（P〉0.05）;在对照组明显升高（P〈0.05）;两组输血后CRP均明显升高,IgA均无明显变化。观察组的输注有效率84.9%,与对照组56.1%相比差异有统计学意义,输血不良反应的发生率也明显减低。对血小板无效输注（PTR）患者进行分析,输血前其血清（IgG、IgM）、补体C3、CIC水平异常者比例较高,PTR患者在输血后体液免疫指标升高更明显（P〈0.05）。结论免疫球蛋白（IgG、IgM）和补体（C3、C4）、CIC、CRP可能参与了血小板无效输注,输注γ辐照机采血小板可以在一定程度上减轻患者的体液免疫反应,提高输注效率。     

An immediate hemolytic reaction induced by repeated administration of oxaliplatin

BACKGROUND: Platinum-based chemotherapy agents have been associated with potentially fatal acute immune-mediated hemolytic anemia. The target antigen, cause of the positive direct antiglobulin test (DAT) and mechanism of hemolysis have been the subject of controversy. CASE REPORT: We report a patient who developed a DAT-positive hemolytic episode after a red cell (RBC) transfusion was delivered during the infusion of her 17th cycle of oxaliplatin. Standard pretransfusion testing was uncomplicated; however, after infusion, the serum was no longer compatible with the transfused units and a strong (4+) panreactive IgG antibody was detected. RESULTS: The patient's serum from 10 days after the episode, only when therapeutic concentrations of oxaliplatin were added, reacted with all RBCs tested using the indirect antiglobulin test (IAT) (3+). The effect was retained with a purified IgG fraction and almost eliminated with IgG-depleted serum. Immunoprecipitation analysis revealed a band with the molecular weight of the Band 3 anion channel only in the presence of the patient's serum and oxaliplatin. CONCLUSION: Our investigations indicated that oxaliplatin interacted with both an IgG antibody and a RBC membrane epitope probably located on the Band 3 anion channel.     

The efficacy of performing red cell elution studies in the pretransfusion testing of patients with positive direct antiglobulin tests

In order to evaluate the efficacy of performing red cell elutions in pretransfusion testing, the serologic records of 638 patients with positive direct antiglobulin tests (DAT) were reviewed. These patients were identified by routine antibody screening procedures that included an autologous control. DAT results on the red cells of these patients showed 279 with IgG and C3d sensitization, 319 with IgG alone, and 40 with C3d sensitization alone. Of 638 patients' red cell eluates, 401 demonstrated no reactivity, 154 demonstrated panagglutination, and 60 demonstrated passively acquired anti-A,B. Only 23 of 638 patients had alloantibody sensitization of their red cells. Of the 23, 19 had serum antibody corresponding to the specificity of antibody detected in the eluate. Thus, only four of 638 (0.6%) eluates gave results unavailable by serum testing alone. This study indicates that routine eluate investigation provides little useful information in assuring compatibility. Serum antibody testing and careful review of the clinical and transfusion history constitute appropriate pretransfusion testing in patients with positive direct antiglobulin tests. Eluate testing should be restricted to cases in which immune hemolysis is suspected clinically.     

Invalidation of antiglobulin tests by a high thermal amplitude cryoglobulin

A multiply transfused patient was referred for evaluation of a transfusion reaction. The direct and indirect antiglobulin tests (DAT, IAT) for alloantibody were negative. However, IgG-coated control cells failed to agglutinate in the negative reactions, casting doubt on their validity. At 4 degrees C, the patient's serum exhibited a large cryoprecipitate (2.9 mg/mL), made up predominantly of an IgG kappa paraprotein and having trace amounts of IgM and C3. Clear serum separated at 37 degrees C became cloudy within 10 minutes at room temperature (RT); within 4 hours, approximately 60 percent of the total precipitable cryoprotein had precipitated. Red cells (RBCs) incubated in fresh serum that had cooled to RT or RBCs obtained from RT or refrigerated samples contained cryoprecipitate that sedimented with the RBCs during washing with RT saline. On resuspension, enough IgG cryoglobulin redissolved to neutralize completely the commercial anti-IgG reagents. If the patient's samples were maintained at 37 degrees C, cryoprecipitate did not form, and RBCs washed four times at 37 degrees C gave valid DAT and IAT reactions. The removal of all cryoprecipitate from the patient's serum by centrifugation after overnight incubation at 4 degrees C also made possible valid antibody screening and compatibility tests.     

Evaluation of the manual hexadimethrine bromide (Polybrene) technique in the investigation of autoimmune hemolytic anemia

The use of the direct manual hexadimethrine bromide (Polybrene) test (DPT) in the investigation of patients for autoimmune hemolytic anemia (AIHA) was evaluated. Seventy-nine blood samples from 68 patients were tested. A direct antiglobulin test (DAT) using monospecific reagents and the DPT were performed, and a concentrated ether eluate was tested. The DAT was positive in 62 (78%) of 79 patients and negative in 17 (22%). There is a good correlation among DAT, eluate, and DPT in demonstrating the presence of immunoglobulin on the red cell surface. In contrast, the DPT does not detect C3d and is often negative in cases of AIHA in which C3d alone is demonstrated by the DAT. In DAT-negative cases, DPT results correlated with reactive eluates. However, in four cases of steroid-responsive, DAT-negative hemolytic anemia, the DPT supported the diagnosis of AIHA when the eluate did not react. The DPT is a useful additional screening test for the investigation of AIHA, but it is not recommended as a replacement for either eluate testing or the DAT.     

Association of positive direct antiglobulin test (DAT) with nonreactive eluate and drug-induced immune hemolytic anemia (DIHA)

Background and Objectives: Drug-induced immune hemolytic anemia is a rare condition that occurs primarily because of drug-induced antibodies, either dependent or independent and positive direct antiglobulin test. Our aim was to evaluate the association of positive DAT with nonreactive eluate and DIHA. Materials and Methods: From 2014–2018, we evaluated 159 patients who presented positive DAT with a nonreactive eluate. Laboratory and clinical analyses were performed including HIV, HBV and HCV testing. All patients were exposed to the following drugs: Dipyrone in 63.5 %, Furosemide in 28.9 %, Metoclopramide in 34.6 % and Ondansetron in 41.5 %. Results: Results of DAT showed IgG in 125 (78.4 %) patients and C3d in 24 (15.1 %) with reactions varying from 1+ to 4+. HIV test was positive in 10 (16.1 %) patients, HBV was positive in 3 (4.7 %) and HCV was positive in, 1 (1.5 %). There was no clinical significance when the parameters of hemoglobin, hematocrit, reticulocytes and LDH were evaluated, only a slight increase in bilirubin, especially, in patients with positive DAT reacting 3+/4+ due to IgG and C3d sensitization. Clinical evaluations showed that all patients were asymptomatic. Conclusions: The association of drugs with positive DAT can be a challenge to transfusion services and immunohematology reference laboratories. There was no evidence of any case of severe hemolysis with clinical repercussion through the clinical and laboratory findings analyzed with the drugs associated with positive DAT. Dipyrone and Furosemide have already been associated with DIHA but there are no studies reporting the association of Metoclopramide and Ondansetron with DIHA.