RHCE(2-9)-D变异产生抗-D引起新生儿溶血病1例

目的 对1例有输血史多次分娩史的RhD阴性孕妇D抗原进行血清学和分子生物学检测,明确RhD表型和基因型。方法 微柱凝胶技术进行母亲和新生儿Rh血型初筛,母亲红细胞经典抗人球蛋白技术进行RhD阴性确认,吸收放散试验确定是否为Del。基因检测确定孕妇RHD基因型。结果 该孕妇结果为Ccdee, RhD确证试验为D阴性,吸收放散试验阴性,基因检测为RHCE(2-9)-D变异,产生抗-D,抗-E。新生儿为CCDee表型,游离抗体检出抗-D和抗-E,放散液检出抗-D。患儿诊断为新生儿溶血病,进行输血、光疗和换血治疗。结论 RHCE(2-9)-D变异型血清学可能表现为D阴性,确证试验阴性,有产生抗-D并引起新生儿溶血病的风险。     

西安地区Rh阴性个体D基因多态性的研究

目的研究西安地区RhD阴性个体的RHD基因多态性分布状况,以及Del血型与其它Rh血型表形之间的关系。方法应用PCR方法检测RhD阴性表型的个体,分析Del血型的Rh表型分布,对特殊样本进行测序分析。结果在607例样本中,检出d/d基因型428例,占70.5%;Del型125例,占20.6%;弱D15型21例,占3.2%;RHD-CE(2—9)-D/d融合基因型31例,占5.0%;DⅥⅢ型2例,占0.4%;Del血型个体中具有C抗原者比例较高,但亦有其它表型存在;鉴定出1例新的RHD无效等位基因。结论对西安地区Rh阴性无偿献血者D基因多态性进行研究,揭示了Rh阴性个体D基因多态性,既有临床意义,也有遗传学意义。     

100例RhD初筛阴性孕妇D放散型表型筛查及基因型分析

目的对100例Rh D阴性孕妇进行D放散型(Del)表型筛查及基因型分析,同时进行抗体筛查和鉴定,以指导Del血型孕妇的产前监测。方法 2016年2月—2016年11月期间,收集100例多次妊娠且初筛为Rh D阴性的孕妇外周血标本,采用抗球蛋白凝胶卡法检测Rh D血型,抗球蛋白凝胶卡法进行直接抗球蛋白试验;采用Rh血型分型卡检测Rh CE抗原;抗球蛋白凝胶卡法进行不规则抗体筛查及抗体鉴定。采用吸收放散试验进行Del血型表型鉴定,同时采用序列特异性聚合酶链反应(PCR-SSP)进行RHD*01EL.01(RHD*1227A)等位基因检测,应用限制性片段长度多态性聚合酶链反应(PCR-RFLP)对Del表型标本RHD基因合子型进行进一步分析;对D变异型标本,进行D抗原表位检测(D-Screen)和RHD基因10个外显子的PCR扩增及直接测序分析。结果 100例标本中检出Del表型27例(27%),均没有产生抗-D,其中21例Rh CE抗原分型为Ccee(77.8%),4例为CCee(14.8%),2例为Cc Ee(7.4%);25例RHD基因型为RHD*01EL.01/01N.01(92.6%),2例RHD*01EL.01/RHD*01EL.01(7.4%)。此外还检出D变异型4例(4%),其中包括2例部分DⅥ3,1例弱D25和1例弱D15,均未产生抗-D。检出Rh D真阴性血型69例,其中产生抗-D 9例(12.7%),且有2例合并抗-C;产生抗-c E合并抗-Jkb1例。结论携带有RHD*01EL.01等位基因的"亚洲型"Del血型孕妇在Rh D阳性胎儿的免疫刺激下,很可能不会产生抗-D,属于完全性Del(complete Del)血型。     

20例输注D~(el)型红细胞的RhD阴性受者的回顾性分析

目的探讨Del型红细胞是否诱导RhD阴性受者产生同种免疫反应。方法用吸收放散试验鉴定Del型血液,对临床输注Del型红细胞的患者进行追踪,采集输血后1周～1年的患者血液标本,RhD抗原初筛阴性的标本采用间接抗球蛋白试验(IAT)进行阴性确认。对患者血清采用凝聚胺法、间接抗球蛋白试验和微柱凝胶法筛查不规则抗体,然后对筛查阳性标本用谱细胞进行不规则抗体鉴定,并进行效价测定。结果采集到输注Del红细胞的RhD阴性受血者样本20例,男性10例,女性10例,女性均有孕产史。Del型受血者5例,男性2例,女性3例,输注Del红细胞后未发生输血反应,不规则抗体筛查阴性。真实RhD阴性受血者15例,男性8例,女性7例,8例男性和5例女性患者输注Del红细胞后未发生输血反应,不规则抗体筛查阴性;2例妊娠女性抗-D为阳性,未出现输血反应。结论 Del型个体未检出抗-D,产生抗-D的个体未检出Del型。Del红细胞导致RhD阴性受者产生同种免疫反应的概率可能很低。     

汉族DEL型孕妇可免去抗-D产前检查

目的鉴于占我国汉族Rh(D)阴性人群约25%的DEL型红细胞具有基本完整的D抗原表位,临床观察汉族DEL型孕妇妊娠Rh(D)阳性胎儿是否产生同种免疫反应。方法跟踪观察和测定207名有妊娠史的Rh(D)阴性孕妇产前和产后血清抗-D,根据RhCcEe表型和PCR检测RHD1227A等位基因鉴别DEL型和真实Rh(D)阴性表型。结果 207名Rh(D)阴性孕妇中,DEL型个体46名(22.2%)均未检测到血清抗-D,即使5名个体有2次或以上生育史,亦未见同种免疫反应。161名真实Rh(D)阴性个体中检出40例抗-D阳性(24.8%);80名有生育史的真实Rh(D)阴性孕妇中30例抗-D阳性(37.5%);20名有2次或以上生育史的真实Rh(D)阴性孕妇中12例存在抗-D(60.0%)。结论 DEL型孕妇妊娠Rh(D)阳性胎儿发生抗-D同种免疫反应的可能很小,我国Rh(D)汉族阴性孕妇中的DEL个体可免去定期的产前抗-D检测,以及免去预防性Rh免疫球蛋白的使用。     

Rh阴性个体输入D~(el)红细胞产生抗-D免疫的研究

目的探讨Rh阴性个体输血中Del型红细胞的D抗原免疫原性。方法对临床输血的Rh阴性患者进行回顾性跟踪检测,采用PCR-SSP法检测标本RHD基因,间接抗球蛋白法检测抗-D效价,流式细胞术对比输血前后抗体强度的变化。结果 2名输注Del型血液的Rh阴性患者,抗-D效价升高,流式细胞术检测结果显示抗-D强度明显升高,说明Del表型红细胞具有一定的免疫原性。结论为了保障临床输血安全,常规血清学检测Rh阴性的供血者,应进一步排除弱D及Del型。     

RhD~(Ⅵ)Ⅲ表型分子生物学研究及其临床意义

目的了解RhD抗原阳性(变异)产生IgG抗-D个体的RHD基因结构。方法采用血清学方法检测个体RhD抗原,鉴定RhD变异个体体内抗体的性质。采用PCR-SSP方法检测其RH血型基因,观察RhD抗原变异体基因构成情况。结果变异个体Rh血型为RhDⅥⅢ型(D抗原不完全型),单倍体型CDe/cde。结论准确的RhD血型鉴定对制定安全有效的临床输血策略和对育龄妇女采取恰当措施预防新生儿溶血病意义重大。     

RhD阴性受血者输注DEL型血液诱发抗-D 1例

目的通过血型血清学试验和分子生物学技术鉴定出1例RhD阴性受血者输注DEL型血液后诱发抗-D产生。方法采用血型血清学方法进行血型抗原及抗体检测,PCR-SSP法进行RhD基因分型检测。结果受血者ABO血型为A型,Rh分型为ccdee,直抗阴性。受血者随机输注的3位供者均为DEL RhD1227A纯合型,且受血者输血后2周抗体筛查为阳性,并鉴定为抗-D。结论 RhD阴性受血者输注DEL RhD1227A纯合型血液后可以引起初次免疫反应,并产生不规则抗-D。所以对于RhD阴性供血者应加强阴性确认,必要时可以增加RhD的基因检测以保障临床输血安全。     

Rh血型系统Del型研究进展

Rh血型系统是人类红细胞血型系统中最为复杂且最具多态性的系统,在临床的重要性仅次于ABO血型系统,而且其分子机理又非常复杂.到目前为止已发现至少49种抗原,其中与临床输血安全密切相关的Rh血型抗原主要有D、C、C、E、e这5种.Rh(D)还存在一些变异体——部分D、弱D和Del等.1984年日本学者Okubo在Rh(D)阴性献血者中首次发现1种非常弱的Rh(D)抗原表达型,命名为Del表型.其非常弱的Rh(D)抗原无法用敏感的间接抗球蛋白试验检出,而只能通过吸收放散试验才可检出.由于人们在Del型是否归于Rh阴性这点上并没有形成共识,故早期Del型献血者血液一直被当作Rh(D)阴性血液而应用于临床输血.     

长春地区献血者RH Del血清学检测

Rh血型D抗原是引起严重新生儿溶血病的主要红细胞抗原.D抗原有多种变异体,如弱D型、部分D型、D放散型(Del)等.Del红细胞膜D抗原非常弱,常规间接抗人球蛋白试验(IAT)测定为阴性,只有通过敏感的吸收放散技术才能检出.Del在不同民族中表型频率差别较大.目前,全国大部分血站提供给临床的RhD阴性血液,都没有检测Del或用PCR方法检测RhD基因及Del基因型,因此对RhDel血液的继续研究就显得由为重要.     

The routine use of Rh-negative reagent red cells for the identification of anti-D and the detection of non-D red cell antibodies

BACKGROUND: Before 1987, fewer than 50 patients per year at the authors' laboratory had a positive antibody detection test due to antepartum Rhesus immunoprophylaxis. However, after 1987, a marked increase was observed in the number of patients who had received Rh immune globulin (RhIG) during pregnancy as part of routine antepartum Rh immunoprophylaxis. In anticipation that an increased use of RhIG during pregnancy would increase the number of patients in whom anti-D was detected by this laboratory, a protocol was developed to abbreviate the process required to identify anti-D. Although this protocol was adopted primarily to address an anticipated increase in antenatal RhIG usage in women, it was also applied to alloimmunized Rh-negative males. STUDY DESIGN AND METHODS: When an Rh-negative patient (male or female) had a reactive screening test for unexpected antibodies and met certain other criteria, the patient's serum was tested with a three-vial set of Rh-negative reagent red cells (Rh-negative screening RBCs), instead of with panels of typed RBCs (panel RBCs), for the identification of anti- D or the detection of non-D antibodies. If the serum under test did not agglutinate or hemolyze Rh-negative screening RBCs, anti-D was identified and no further testing was performed. If the serum agglutinated or hemolyzed Rh-negative screening RBCs, conventional testing with panel RBCs was done to determine the antibody specificity. RESULTS: Rh-negative patients (n = 1174) who had reactive screening tests for unexpected antibodies were tested with Rh-negative screening RBCs; 1079 were found to have anti-D as a single antibody. Seven of these patients subsequently developed a non-D alloantibody, after transfusion or pregnancy, and one patient had anti-C that escaped detection at the time of initial testing with Rh-negative RBCs (a false- negative result). Ninety-two patients had anti-D in combination with a non-D antibody, and three patients had a non-D antibody but not anti-D. Use of the anti-D identification protocol actually reduced the laboratory workload by 176 College of American Pathologists workload units per month, in spite of a marked increase in the number of patients in whom anti-D was detected. No hemolytic transfusion reaction was attributed to the abbreviation of anti-D identification. CONCLUSION: The identification of anti-D may be abbreviated without jeopardizing patient safety. Such a protocol can reduce laboratory workload and might be particularly appealing to health care facilities that perform antibody detection testing on large numbers of Rh-negative pregnant women, especially if antepartum RhIG is administered routinely.     

Fetomaternal hemorrhage: incidence, risk factors, time of occurrence, and clinical effects

Most women have only very small amounts of fetal blood in their circulations following pregnancy and delivery: the volume is less than 0.5 mL of whole blood in 93 percent of women, less than 1 mL in 96 percent, and less than 2 mL in 98 percent. FMH of 30 mL or more occurs in just 3 of 1000 women. When the FMH was 150 mL or more, 15 of 41 infants did not survive Rh-negative women with FMH of more than 30 mL of Rh-positive whole blood are at increased risk of Rh immunization, and thus the outcome of their future pregnancies also may be affected. ABO-compatible fetal red cells that have entered the maternal circulation have a life span similar to that of adult cells. ABO-incompatible fetal red cells may be cleared rapidly, but in some cases they circulate for weeks. Most FMHs of 30 mL or more occur before labor, delivery, or cesarean section. The majority occur with minimal clinical signs and symptoms in apparently normal pregnancies. The identification of postpartum Rh-negative women who have 30 mL or more of Rh-positive fetal blood in their circulation is important so that sufficient RhIG for immune suppression can be administered. It appears that more than one-half of women with FMH of 30 mL or more would not be identified if protocols were adopted to test only women in pregnancies considered to be at high risk.     

Rh阴性血型患者输血相关问题探讨

Rh血型系统是重要的红细胞血型系统,目前临床输血治疗对于Rh阴性血液的需求仍处于紧张状态.实际临床工作中,通过典型案例分析总结针对Rh阴性血型患者输血的临床经验,建立规范化的血库管理模式,分析Rh阴性血型患者的输血类型,并且积极普及Rh阴性血型的相关知识,尽力减少临床输血过程中导致的输血反应等问题,从而达到安全、合理输血的目的.     

Studies on Rh Prophylaxis

One hundred and seventy-eight (178) Rh-negative volunteers, distributed into a treated and control series of six groups each, were studied to establish: (i) approximately 70 per cent of Rh-negative individuals are susceptible to being immunized by a single injection of Rh-positive blood; (ii) that, for immunologically susceptible individuals, the frequency of immunization increases with the volume of Rh-positive erythrocytes administered; and (iii) that a possible relationship exists between potency of Rh immune globulin and effectivity. This relationship can be used to calculate an effective dose of Rh immune globulin in the treatment of large feto-maternal hemorrhages or accidental tranfusions of Rh-positive blood to Rh-negative women.     

RHD711delC等位基因检测和频率计算

目的了解RHD711delC等位基因或RHD710delC在一般人群中的基因频率。方法对890 403名汉族无偿献血者,采用盐水法和间接抗人球蛋白试验（IAT）测定D抗原,PCR-SSP检测RHD711delC等位基因以及RHD合子型,应用遗传统计方法分析基因频率。结果 Rh（D）表型检测显示2 385人为Rh阴性,108人为D抗原弱阳性表型（包括弱D型和部分D型）,其余887 910人为Rh阳性。2 385名Rh阴性个体中检测到19人携带RHD711delC等位基因（0.80%）,均为RHD711delC/d杂合子;108名D抗原弱阳性个体中,未检出RHD711delC携带者;随机抽检960名Rh阳性样本,发现1人为RHD711delC/RHD＋。根据上述结果,计算RHD711delC在被检人群中的基因频率为0.000 530;根据RHD711delC在Rh阴性人群的检出率,推算其在一般人群中的基因频率为0.000 225。结论 RHD711delC等位基因在中国汉族人群中的等位基因频率在0.000 225～0.000 530之间。     

The risk assessment study for hemolytic disease of the fetus and newborn in a University Hospital in Turkey

Maternal red-cell alloimmunization occurs when a woman’s immune system is sensitized to foreign red-blood cell surface antigens, leading to the production of alloantibodies. The resulting antibodies often cross the placenta during pregnancies in sensitized women and, if the fetus is positive for red-blood-cell surface antigens, this will lead to hemolysis of fetal red-blood cells and anemia. The most severe cases of hemolytic disease in the fetus and newborn baby are caused by anti-D, anti-c, anti-E and anti-K antibodies. There are limited data available on immunization rates in pregnant women from Turkey. The aim of the present study was to provide data on the frequency and nature of maternal RBC alloimmunization in pregnant women in a tertiary care hospital. In this study, we retrospectively evaluated the indirect antiglobulin test results of Rh-negative pregnant women performed in our Blood Bank between 2006 and 2012. Indirect antiglobulin test positive women also underwent confirmatory antibody screening and identification. During the study period, 4840 women admitted to our antenatal clinics. With regards to the major blood group systems (ABO and Rh), the most common phenotype was O positive (38.67%). There were 4097 D-antigen-positive women (84.65%) and 743 women with D-antigen-negative phenotype (15.35%). The prevalence of alloimmunization was found to be 8.74% in D-antigen negative group. Despite prophylactic use of Rh immunglobulins, anti-D is still a common antibody identified as the major cause of alloimmunization in our study (anti-D antibody 68.57%, non-D antibody 31.42%). While alloimmunization rate to D antigen was 6.46%, non-D alloimmunization rate was 2.69% among Rh-negative pregnant women. Moreover, detailed identification facilities for antibodies other than anti-D are not available in most of centers across Turkey. However, large-scale studies on pregnant women need to be done in order to collect sufficient evidence to formulate guidelines and to define indications for alloantibody screening and identification.     

Overestimation of fetomaternal haemorrhage by the acid-elution technique in mothers with β-thalassaemia minor

Summary. In Rh-negative women, it is important to quantify the magnitude of an Rh-positive fetomaternal haemorrhage (FMH) so that sufficient Rh immune globulin (RhIg) can be administered early in the postpartum period to prevent alloimmunization. The standard post-partum dose of Rhlg varies from 100 μg in the UK to 300 μg in North America. It is therefore important to identify all Rh-negative women who have had an FMH greater than 10 ml in the UK or greater than 30 ml in North America because an FMH greater than these amounts will affect the dose of RhIg that is administered. As acid-elution techniques can overestimate the magnitude of an FMH in the presence of an elevated maternal haemoglobin F level, we performed a prospective study to determine how often this occurred. Of 1,894 consecutive Rh-negative mothers who delivered Rh-positive infants, whose blood was screened for an FMH greater than 10 ml of fetal blood using an acid-elution procedure, 11 were found to have an FMH over 10 ml. In five of these 11 women, the volume of FMH was less than 10 ml using an alternative technique (rosette test) to assess the FMH size. Six of these women were found to have β-thalassaemia minor on the basis of a low MCV, and high haemoglobin A₂ and/or high haemoglobin F levels. In five of these the FMH was significantly overestimated by the acid-elution technique compared to the rosette technique. Therefore, in the presence of a maternal condition, which may result in an elevated haemoglobin F level, an FMH estimated to be over 10 ml in the UK or 30 ml in North America using an acid-elution procedure, should be confirmed by an alternative technique, which does not involve the estimation, directly or indirectly, of haemoglobin F.     

Unusual amniotic fluid spectral analysis in Rh sensitization.

In our experience delta 450 millimicron OD measurements have shown a progressive decline during the latter half of gestation in pregnancies unaffected by fetal hemolytic disease. However, we have seen two Rh-negative patients in whom there was a rise in delta 450 millimicron OD between 24 and 27 weeks' gestation and who subsequently bore Rh-negative infants. In these two cases the increase between 24 and 27 weeks was from approximately 0.140 to about 0.200. Both infants were unaffected with fetal hemolytic disease. Therefore, we recommend caution in interpreting modest increases in delta 450 millimicron OD during this period of gestation.     

Transient Production of Anti-LW by LW-positive People

As the result of pregnancy immunization two Rh-negative women produced anti-D and anti-LW, while an Rh-positive woman produced anti-LW in response to transfusion. The first two were both initially LW-negative and direct antiglobulin test negative. One then became direct antiglobulin test positive, and this lasted as long as anti-LW was present in her plasma. Other observations suggest this may not be a rare occurrence.