新生儿溶血病Rh血型免疫性抗体检测的分析

目的:探讨母婴Rh血型不合免疫性抗体的特异性及其效价对新生儿溶血病(HDN)的影响。方法:采用血型血清学检测技术对母婴进行ABO及Rh血型鉴定,对15例母婴Rh血型不合HDN患儿的血标本进行直接抗球蛋白试验、抗体游离试验和抗体放散试验,采用间接抗球蛋白试验对患儿及其母亲的血清进行ABO以外血型不规则抗体筛选、特异性鉴定及效价测定。结果:在15例Rh HDN患儿中检出抗-D 4例(26.7%),抗-E 6例(40.0%),抗-cE 3例(20.0%),抗-Ce 2例(13.3%);Rh血型免疫性IgG抗体效价为1∶8～1∶128。结论:产前对孕妇夫妇进行ABO、Rh血型鉴定及Rh血型免疫性抗体筛查及特异性鉴定,产后对患儿及时进行检测诊断和治疗,对减少HDN患儿的受害程度和保证优生优育均有重要的临床意义。     

154例新生儿ABO溶血病的检测结果分析

目的了解母婴ABO血型不合的新生儿溶血病(HDN)的发病率,以及HDN血清学检测结果与新生儿出生后天数的关系。方法母婴血型鉴定,新生儿溶血病三项试验,即直接抗人球蛋白试验,游离抗体试验,抗体释放试验。结果对我院出生或治疗的154例有不同程度黄染或临床疑似为新生儿溶血病的新生儿进行了血清学检测。结果新生儿溶血病的发病率为54.5%(84/154)。其中出生后0~2d组的阳性率为90%(27/30),出生后3~4d组的阳性率为69.7%(23/33),出生后5~7d组的阳性率为47.1%(24/51),出生后8~15d组的阳性率为25%(10/40)。结论新生儿溶血病的发病率较高,其中出生后0~2d组的检测阳性率较高于其它组。     

新生儿溶血病ABO血型免疫性抗体检测分析

目的:探讨血型免疫性IgG抗体对母婴ABO血型不合新生儿溶血病的影响.方法:采用抗人球蛋白法、微柱凝胶法对临床有新生儿高胆红素血症的患儿进行血型血清学检测,对母婴ABO血型不合的患儿血标本进行直接抗人球蛋白试验、抗体游离试验和抗体放散试验,检测免疫性IgG抗体的特异性.结果:在476例临床有新生儿高胆红素血症的患儿中,由母婴ABO血型不合引起的新生儿溶血病为59.5%(283/476).其中直接抗人球蛋白试验阳性率为31.4%(89/283),抗体游离试验阳性率为79.5%(225/283),抗体放散试验阳性率为100%(283/283);在283例ABO新生儿溶血病中,由IgG抗A引起者占48.1%(136/283),由IgG抗B引起者占51.9%(147/283).结论:ABO血型为A型或B型的分布与新生儿溶血病的发病率无显著性差异.     

105例母婴ABO血型不合的新生儿中溶血病的实验室检测结果分析

目的了解母婴ABO血型不合的新生儿中新生儿溶血病(HDN)的发病率,并为临床提供HDN的诊断和防治的依据。方法夫妻及新生儿血型鉴定;对新生儿血做直接抗人球蛋白试验、抗体释放试验、游离抗体试验;对产后母血做IgG抗A(B)抗体效价试验以及ABO以外抗体试验。结果对我院出生的105例不同程度黄染的新生儿进行新生儿溶血病的实验室检测,结果新生儿溶血病的总发病率为22.9%(24/105)。其中母夫婴O-A-A型血组的阳性率是33.3%(12/36),O-B-B型血组的阳性率为13.3%(6/45),O-AB-A型血组的阳性率是28.6%(4/14),O-AB-B型血组的阳性率是20.0%(2/10).产后母亲血清IgG抗A(B)抗体效价大于1∶32者占71.4%。结论新生儿溶血病的发病率较高,其中O-A-A型血组的发病率较高于其它组;母亲血清IgG抗A(B)抗体阳性率很高。我们建议:凡以往不知原因的死胎、流产、输血史或新生儿重度黄疸史的孕妇,对产前ABOHDN效价高的母亲、产后黄疸持续不退的婴儿及时做新生儿溶血病的检测,做到早发现早防治。     

新生儿溶血病红细胞血型免疫性抗体的特异性分析

目的:观察母婴血型不合新生儿溶血病(HDN)红细胞血型免疫性抗体的检出率及其特异性.方法:采用试管法和微柱凝胶法对母婴血标本进行ABO、Rh血型鉴定及不规则抗体筛查,对有黄疸症状的新生儿血标本进行直接抗人球蛋白试验、抗体游离试验、抗体放散试验、抗体特异性鉴定及其效价测定.结果:在298例由红细胞血型免疫性抗体引起的HDN患儿中,检出抗A 62例(20.8％),抗B 65例(21.8％),抗A+抗AB 87例(29.2％),抗B+抗AB 6例(2.0％);抗M4例(1.3％),抗N1例(0.3％);抗D4例(1.3％),抗E7例(2.3％),抗cE 3例(1.0％).由ABO及MN血型免疫性抗体引起HDN者,直接抗人球蛋白试验多为弱阳性,由Rh血型免疫性抗体引起HDN者,直接抗人球蛋白试验多为强阳性;微柱凝胶法的凝集强度高于试管法;HDN患儿出生后24 h内阳性检出率为91.5％.结论:HDN血型免疫性抗体的特异性主要为ABO系统的抗A、抗B、抗AB,其次为Rh系统的抗E、抗D、抗cE及MN系统的抗M、抗N.     

41例新生儿溶血病患者血型血清学检测结果分析

目的：探讨不同的免疫性血型抗体与新生儿溶血病的关系,为新生儿溶血病提供诊断依据。方法采用微柱凝胶技术对41例新生儿溶血病的患者进行ABO血型、Rh(D)血型、直接抗人球蛋白、游离、放散和不规则抗体筛查及抗体鉴定试验,确定患者体内是否有免疫性IgG抗体及IgG抗体特异性。结果41例新生儿溶血病患者中,ABO血型不合者38例,占92.68%,其中由免疫性IgG抗原A抗体引起者21例,由免疫性IgG抗原B抗体引起者17例。Rh血型不合者3例,占7.32%,其中2例由抗-D引起,1例由抗-E引起。41例新生儿溶血病的患者中,直接抗人球蛋白试验阳性者20例,阳性率为48.78%,游离试验阳性者36例,阳性率为87.80%,放散试验阳性者41例,阳性率为100%。结论母婴血型不合的新生儿溶血病主要发生于ABO血型系统,以母亲为O型,患者为A型或B型最为常见;放散试验对新生儿溶血病的诊断最有价值。     

西安地区孕妇产前新生儿溶血病(HDN)筛查检测结果分析

目的了解西安地区孕妇中ABO血型不合及Rh血型不合的发生率,为临床提供孕妇HDN筛查的实验室依据.方法夫妻双方的ABO血型鉴定、RhD血型鉴定、孕妇不全抗体检查(抗体筛选,抗体鉴定,效价测定).结果孕妇ABO血型不合发生率为38.26%(1065/2783),ABO血型不合与Rh血型不合同时存在者为64.70%(11/17),Rh血型不合发生率为50.00%(2/4).结论孕前夫妻双方血型鉴定,孕早期HDN筛查对于预测胎儿与新生儿患免疫性溶血病可能性,以便及时采取相应措施进行预防,是十分必要的.     

IgGA(B)抗体与新生儿溶血病关系的探讨

目的探讨IgGA(B)抗体与新生儿溶血病的关系。方法对2693例新生儿黄疸症状的患儿作ABO、Rh(D)血型检查,直接抗人球蛋白试验、游离抗体测定及放散试验。结果在2693例患儿中母婴ABO血型不合所致新生儿溶血病有334例,母婴O/A所致的HDN为161例,O/B所致的HDN为171例,A/ABHDN1例,B/ABHDN1例。结论对新生儿黄疸患儿及时进行ABOHDN血型血清学检测,有助于早期诊断及时诊治。     

新生儿黄疸患儿血清学检测结果分析

目的了解新生儿黄疸患儿中溶血病（HDN）发病情况。方法对526例新生儿黄疸患儿采用盐水法检测ABO、Rh血型,采用直接抗人球蛋白试验（DAT）、游离抗体试验、抗体释放试验检测,分析其结果阳性率和临床符合率,并进行统计学分析。结果诊断为新生儿溶血病180例,其中ABO血型系统不合的169例,Rh血型系统不合的11例;新生儿黄疸血清学检测阳性率及在HDN诊断中的临床符合率分别为22例（4.2%）和22/180例（12.2%）、368例（70.0%）和368/180例（204.4%）、180例（34.2%）和100.0%,三者间比较均有显著差异（P〈0.05）。结论血型血清学检测对新生儿溶血病的诊断具有重要的临床价值;ABO-HDN的患儿DAT结果大多数是阴性,游离抗体试验在HDN诊断中假阳性发生率较高,只能作为参考而不能作为诊断依据,抗体释放试验是诊断ABO-HDN最灵敏、最可靠的方法。     

母婴血型不合HDN患儿致敏红细胞的抗体特异性

目的:调查母婴血型不合新生儿溶血病(HDN)患儿致敏红细胞的抗体特异性,比较不同抗体致敏新生儿红细胞所致HDN的血型血清学特征.方法:对黄疸新生儿,鉴定母婴血型,做新生儿红细胞直接抗球蛋白试验(DAT),检测母婴血浆及新生儿红细胞放散液中可致敏新生儿红细胞的血型抗体以诊断HDN.结果:血型血清学诊断为HDN的252例患儿中,致敏红细胞的抗体分别为:抗-A 40例、抗-B 40例、抗-A 抗-AB 92例、抗-B 抗-AB 65例、抗-AB 4例、抗-A 抗-M 1例、抗-M 3例、抗-c 1例、抗-cE 1例、抗-E3例、抗-D 2例;由ABO血型抗体及抗-M所致HDN者DAT多为阴性或弱阳性,由Rh血型抗体致HDN者DAT均为强阳性,ABO、Rh血型抗体及抗-M致HDN患儿红细胞热放散液中致敏红细胞的抗体效价均高于血浆游离抗体.结论:被调查的HDN患儿绝大多数由来自O型母亲的IgG抗-A、抗-B及抗-AB所致,其次为抗-M及抗-E,由抗-D引起的HDN呈逐步减少的趋势.     

母婴ABO合并Rh-ccdee/ccDEe血型不合HDN实验诊断与鉴别诊断的研究

目的：探讨母婴ABO合并Rh血型不合和Rh两座位3位点表型不合HDN实验诊断与鉴别诊断,为临床早期防治提供实验依据。方法：用吸收放散和直抗作为实验诊断,用A-c、B-c和O-c阴性红细胞鉴别诊断,O型Rh-ccdEe、O型Rh-CCDee及A型Rh-CCdee组合谱细胞鉴别两座位多位点表型不合Rh-HDN,盐水法鉴定抗体性质。结果：①母婴ABO/Rh血型：母婴1血型鉴定为 O-A及Rh-ccdee/ccDEe交叉不合、母婴2鉴定为A-A及Rh-ccdee/CcDEe两座位3位点表型不合;②抗体筛选：母婴1不规则抗体均为强阳性（4+）;母2为不规则抗体强阳性（4+）、婴儿2为阳性（+）。③抗体鉴定与鉴别诊断：母1为 IgG抗A合并IgG抗D,IgG抗E缺失,IgG抗A效价 256,IgG抗D效价128;母2为IgM抗E合并IgG抗D,IgM抗C缺失,IgG为抗D效价128,IgG抗E及抗C缺失。④鉴别诊断：婴儿1证实为IgG抗A合并IgG抗D-HDN;婴儿2 证实为Rh抗D-HDN,IgG抗E及抗C缺失。结论：用A-c、B-c、O-c阴性红细胞和Rh单特异性组合谱细胞,可以鉴别诊断ABO合并Rh HDN以及Rh单一性或混合性HDN。     

微柱凝胶技术应用于新生儿溶血病的实验室诊断

目的采用微柱凝胶技术进行新生儿溶血病（HDN）的实验室诊断分析。方法对1627例新生儿进行血型血清学分析,包括母婴血型、直接抗人球蛋白试验、血清游离抗体试验、热放散试验、ABO血型系统以外的抗体鉴定试验。结果 1627例病例中确诊为HDN的有163例,其中母-婴血型为O-A及O-B的最多,共占97.55%。确诊为HDN血清学结果中放散试验和游离抗体试验同时阳性最多,占76.07%。诊断为HDN可疑病例血清学结果中,直接抗人球蛋白试验阳性率最多,占79.55%。在出生后3～7d内确诊HND的患儿数最多,占66.26%。ABO血型系统以外的抗体鉴定试验阳性有6例,其中5例诊断为RhHDN。结论红细胞抗体释放试验、直接抗人球蛋白试验和血清游离抗体试验是早期诊断HDN的有效方法。HDN检出率与所采用的实验方法和采血时机有很大关系。     

Prenatal screening

In most countries, pregnant women are early in pregnancy typed for ABO and RhD and screened for the presence of red cell antibodies. Maternal red cell alloantibodies of IgG class are actively transported by the placenta to the fetus and destruct the fetal red cells, if these carry the involved antigen leading to haemolytic disease of the fetus and newborn, HDFN. HDFN is a disease which –if untreated– can cause perinatal mortality and morbidity, with a substantial risk for long-term sequelae. In most cases of HDFN, alloimmunization against the RhD antigen is involved, although introduction of anti-D immunoglobulin (Ig) prophylaxis drastically decreased anti-D-mediated HDFN. In the last decade, non-invasive fetal genotyping, with cell-free fetal DNA present in maternal plasma, for RhD, RhC, Rhc, RhE and K became a diagnostic reality. In a few countries non-invasive fetal RhD genotyping has already been implemented to target antenatal and postnatal anti-D Ig prophylaxis to RhD-negative women carrying RhD-positive fetuses. Both applications of non-invasive fetal genotyping are summarized in this article.     

Direct enzyme linked antiglobulin tests (ELAT) for detecting in-vivo sensitized erythrocytes: evaluation of screening for ABO incompatibility of newborn

ABO incompatibility of the newborn is one instance where immune hemolysis may present with a negative direct antiglobulin test (DAT) and therefore a simple sensitive test for detecting sensitization would be useful in this clinical situation. To evaluate the usefulness of ELAT in detecting in-vivo sensitized red cells, 1608 maternal-baby pairs were screened for ABO incompatibility over a period of 10 mth. Of 251 ABO-incompatible babies, there were 49 (19.5%) with positive DAT, but an additional 67 (26.7%) were ELAT positive. These were eluate positive as well, indicating that the increased number with sensitized cells as shown by ELAT is due to detection of in-vivo sensitized cells. The positive predictive value for ABO hemolytic disease of the newborn (HDN) is 48%, which is two times that of DAT. Calculating the difference of the absorbance from baseline (delta OD) may give an indication of degree of sensitization which together with the maternal antibody titre would aid us in the estimation of antigen dosage on the baby's red cells and in the appraisal of the role of antigen dosage in HDN.     

After 70 years,serological RhD determination remains a challenge: DNA to the rescue

After ABO system antigens, RhD is the most clinically significant blood group antigen. This is reflected in its high immunogenicity and potential to cause haemolytic transfusion reactions (HTR) and severe haemolytic disease of the newborn (HDN). Thus, correct determination of the RhD antigen is essential for a safe transfusion strategy and adequate indications of anti‐D immunoglobulin prophylactic administration. The RhD determination challenge started in 1939 with a case history of fatal HDN and HTR in a mother who was transfused with her husband′s blood. Subsequent findings of an antibody reacting with 80% ABO compatible red blood cells (RBCs) in the serum of the afflicted woman lead to the hypothesis that the mother was lacking an antigen present on the father's and fetal RBCs and that her production of a corresponding antibody was responsible for both HDN and HTR. This hypothesis was proven to be correct. The following year, the effort to determine the origin of a causal antigen coincided with an animal immunization experiment where a similarly reacting antibody developed by injecting Rhesus monkey RBCs into rabbits and guinea‐pigs, as well as with another publication of HTRs after ABO compatible blood transfusions in patients with antibodies of apparently identical specificity. This first challenge in the ‘pre‐DNA’ era was slightly erroneously named in humans as the anti‐Rhesus antibody, and 20 years later, the animal antibody was designated anti‐LW. Meanwhile, the complexity of Rh group of antigens increased and several genetic models were suggested to explain it: Rh‐Hr single gene, three genes C, D and E and finally a two locus model (which a few years later was shown by molecular analysis to be the accurate one). At the beginning, serological determination of RhD was performed using human anti‐D, first by direct agglutination (IgM), later also using the Coombs’ test (IgG). Weak D antigens were a challenge at the time: the term ‘D^U’ (now obsolete) was used for those antigens negative in direct agglutination and positive in the Coombs’ test. Weakening of the D antigen could be caused indirectly by the ‘position effect C in Trans’ or directly (mutations in the RHD gene, usually point mutations coding for amino acids in transmembranous and intracellular parts of the polypeptide). Extremely weakened D antigens are called D_el, and these are serologically detected only by adsorption/elution tests (multiple genetic mechanisms). The next challenge came in the 1960s, with the observation of qualitative D variants – partial D antigens. The first cases were detected after the development of allo‐anti‐D in D‐positive individuals – immunized against that part of the D protein missing in their D epitope mosaic. Mutual reactivity of anti‐D from these individuals with different partial D antigens established the basis for their classification. The increase in partial D types accelerated dramatically after the development of the mouse hybridoma technique and the production of numerous different monoclonal anti‐Ds. According to patterns of reactivity of different partial Ds after two international workshops, the number of D epitopes reached 30. DNA techniques subsequently helped refine discrimination between partial D types. The number of described D antigen variants will surely rise after this ISBT congress and Working Party meeting. A further increase can be anticipated once NGS will provide new information and more data will come from large Asian and African populations and the polymorphic complexity of RhD and the whole Rh system will expand.     

新生儿Rh溶血病的检查分析与晚期贫血

目的对11例Rh新生儿溶血病血清抗体进行分析。方法采用直接抗人球蛋白试验、游离抗体试验、放散试验检测ABO以外的抗体，采用微柱凝胶间接抗人球蛋白试验进行抗体鉴定。结果11例新生儿Rh溶血病中检出抗-D7例，抗-E3’例，抗-E、C1例，其中1例抗-E新生儿溶血病患儿发生了晚期贫血。结论根据不规则抗体的类型，及时为新生儿溶血病患儿选择相合的血液进行换血和综合治疗，其疗效显著。同时提醒大家要注意新生儿溶血病患儿有发生晚期贫血的可能。     

母儿ABO血型不合孕妇血清IgG抗体效价与新生儿溶血的相关性分析

目的探讨母亲孕期血型血清学的IgG抗体效价与新生儿ABO溶血病(HDN)的关系。方法选取2005年1月～2009年12月在我院分娩的2168例ABO血型不合孕妇及其新生儿为研究对象。以分娩前孕妇最后一次IgG抗A(B)效价值为标准,效价≥1∶64者列为研究范围。新生儿HDN则观察溶血3项筛查指标及其红细胞、血红蛋白、网织红细胞等。结果 2168例ABO血型不合孕妇中血清IgG抗体效价1∶64有710例,1∶128有800例,1∶256有519例,效价≥1∶512有171例。HDN发病患儿共529例。2次及以上妊娠孕妇的抗A、抗B抗体效价值大于1次妊娠者,差异有显著性,P<0.05;年龄30-40岁组孕妇的抗A和抗B抗体效价高于21-29岁组,差异有显著性,P<0.05;母亲IgG抗A或抗B抗体效价与新生儿ABO溶血的发生率相关,r=0.8119,P<0.05。结论孕妇血清中血型免疫性抗体IgG是引起HDN的主要原因,且随着IgG抗体效价的增高,HDN的发病率也增高,但不是HDN发病的唯一因素,妊娠次数的增多、年龄增大、不良生育史等因素可加重IgG抗体效价对HDN的发生。     

Severe hemolytic disease of the newborn in a group B African-American infant delivered by a group O mother

Maternal-fetal ABO incompatibility is a common hematological problem affecting the newborn. In general, hemolysis is minimal and the clinical course is relatively benign, rarely causing the escalating levels of hyperbilirubinemia and significant anemia commonly associated with Rh hemolytic disease of the newborn (HDN). The incidence of HDN ranges from one in 150 births to 1:3000 births, depending on the degree of anemia and level of serum bilirubin. The etiology of ABO hemolytic disease of the newborn (ABO-HDN) is complex because anti-A and anti-B antibodies are composed mainly of IgM. Since only IgG antibodies cross the placenta, those pregnant women with high levels of IgG anti-A,B, anti-A, or anti-B with an ABO incompatible fetus will be the ones to give birth to an infant with ABO-HDN. We describe a case of a B/Rh positive term newborn born to an O/Rh negative African-American mother demonstrating aggressive hemolysis and a robust response of the bone marrow. This case was successfully managed with phototherapy and simple RBC transfusion without the need for exchange transfusion.     

中国新疆汉族和维吾尔族Rh阴性表型的分布特征研究

目的 对比研究新疆汉族和维吾尔族Rh阴性表型的分布特征.方法 选取新疆医科大学第一附属医院住院治疗的Rh阴性患者样本,其中汉族有效样本91例和维吾尔族有效样本231例.分别采用Anti-C、Anti-c、Anti-E、Anti-e单克隆抗体鉴定Rh阴性表型,及采用人类红细胞RhD基因分型试剂盒(筛查型)(PCR-SSP法)测定Rh阴性的基因分型.结果 新疆汉族和维吾尔族Rh阴性人群中以ccee表型最为常见,且维吾尔族ccee表型频率高于汉族(P<0.05).在RhC/c表型分布上,两族均大部分为cc表型,维吾尔族人群cc表型频率高于汉族,Cc表型频率低于汉族(P<0.05).在RhE/e表型分布上,两族均大部分为ee表型,而维吾尔族人群ee表型频率高于汉族,Ee表型频率低于汉族(P<0.05),均存在统计学差异.在Rh阴性的基因分布上,两族均以d/d最为多见,维吾尔族d/d远远高于汉族(90.58 %比45.05 %),而汉族的DEL型高于维吾尔族(19.78 %比1.35 %),两组比较(P<0.05),差异具有统计学意义.结论 新疆维吾尔族人群Rh阴性表型分布特征与汉族人群相比既有相似之处,又有其自身分布特点,可为后续的血型遗传学及血型基因进化研究提供参考,临床RhD抗体导致的新生儿溶血病的防治提供参考依据.     

A fatal case of severe hemolytic disease of newborn associated with anti-Jk(b)

The Kidd blood group is clinically significant since the Jk antibodies can cause acute and delayed transfusion reactions as well as hemolytic disease of newborn (HDN). In general, HDN due to anti-Jk(b) incompatibility is rare and it usually displays mild clinical symptoms with a favorable prognosis. Yet, we apparently experienced the second case of HDN due to anti-Jk(b) with severe clinical symptoms and a fatal outcome. A female patient having the AB, Rh(D)-positive blood type was admitted for jaundice on the fourth day after birth. At the time of admission, the patient was lethargic and exhibited high pitched crying. The laboratory data indicated a hemoglobin value of 11.4 mg/dL, a reticulocyte count of 14.9% and a total bilirubin of 46.1 mg/dL, a direct bilirubin of 1.1 mg/dL and a strong positive result (+++) on the direct Coomb's test. As a result of the identification of irregular antibody from the maternal serum, anti-Jk(b) was detected, which was also found in the eluate made from infant's blood. Despite the aggressive treatment with exchange transfusion and intensive phototherapy, the patient died of intractable seizure and acute renal failure on the fourth day of admission. Therefore, pediatricians should be aware of the clinical courses of hemolytic jaundice due to anti-Jk(b), and they should be ready to treat this disease with active therapeutic interventions.