流式细胞术检测不同RhD血清型红细胞RhD抗原的表达

本研究旨在建立流式细胞术(FCM)检测红细胞表面RhD抗原的实验方法,并研究不同RhD血清型红细胞表面D抗原表达的差异。选取标准的RhD( )和RhD(-)细胞按1∶1比例混合,用间接标记法[一抗为IgG抗-D,二抗为FITC-抗-IgG F(ab′)2]染色,应用FCM检测RhD( )细胞的比例,并确立IgG抗-D的最佳使用稀释度。采用FCM检测RhD阳性、弱D型、RhDel型、RhD阴性红细胞表面D抗原数量。结果显示,IgG抗-D单克隆抗体的最佳使用稀释度是1∶4,1×106/50μl。RhD阳性、弱D型、RhDel型、RhD阴性者RhD( )红细胞百分率分别为(96.8±2.97)%、(79.5±9.88)%、(47.8±11.43)%、(3.7±2.96)%,红细胞表面D抗原平均荧光强度依次为33.3±6.21道、18.6±5.39道、7.10±1.17道、0.79±0.55道。结论:4种不同血清表现型的RhD抗原数量的差异有显著性,以RhD阳性的抗原数量最多,弱D型次之,RhDel型最少。     

流式细胞术检测RhD抗原及其临床应用

目的　探讨流式细胞术 (FCM)检测RhD抗原及其临床应用价值。方法　选取标准的RhD(- )和RhD(+)细胞按 9个不同比例混合 ,分别用直标法 (FITC 抗 D)和间标法 (抗 D和FITC 抗 IgG)染色 ,在FCM上作细胞相对和绝对计数。选择合适检测条件 ,对 2名RhD(- )患者误输RhD(+)红细胞后作RhD(+)细胞计数。结果　与间标法相比 ,直标法检测结果更接近理论值。而单平台和双平台计数法与理论值间无显著性差异 (F =0 .0 32 5 ,P >0 .0 5 )。对 2名RhD(- )的 3份不同时间样本检测发现 ,RhD(+)细胞相对计数分别为 4 .73%、1 0 .87%和 2 0 .86 % ,绝对计数分别为 0 .1 2 4× 1 0 1 2 /L、0 .2 4 5× 1 0 1 2 /L和 0 .5 1 7× 1 0 1 2 /L。结论　FCM能检测RhD(- )细胞中不同浓度的RhD(+)细胞 ,可应用于临床不合血液输注后嵌合态异体细胞的检测     

RhD(-)孕产妇的RhD同种免疫分析及围产期孕妇新生儿溶血病的监测、预防和治疗

目的分析304名RhD(-)孕产妇RhD同种免疫发生情况,探讨RhD(-)孕产妇抗-D产生的影响因素,建立正确的围产期孕妇RhD新生儿溶血病的监测、预防和治疗方案。方法采用标准血清学方法对3975名孕产妇及其丈夫进行ABO及RhD抗原鉴定。对RhD抗原鉴定为阴性的标本,进一步采用间接抗球蛋白方法检测RhD抗原,以排除或确认弱D型或部分D表型。对所有RhD(-)孕产妇及其丈夫进行CcEe表型的血清学分型。采用抗球蛋白方法对所有RhD(-)孕产妇标本进行不规则抗体初筛,对初筛阳性者进一步用鉴定细胞做抗体鉴定及抗体效价测定并采用PCR-SSP方法确定是否为DEL型。结果 3975份孕产妇标本中304份经鉴定为RhD(-),其中29份产生了抗-D,本组调查中RhD(-)孕产妇抗-D产生的比例为9.54%。29名产生抗-D的RhD(-)孕产妇中,夫妇ABO血型相合者24名(82.76%),不合者5名(17.24%)。经分子生物学方法鉴定,29名产生抗-D的Rh(-)孕产妇均排除DEL表型。结论 RhD孕产妇RhD同种免疫的发生受多种因素的影响,DEL型孕产妇产生抗-D的几率较低。应及时、定期监测RhD围产期孕妇的抗-D水平,对未产生抗-D的孕妇应给予抗-D免疫球蛋白治疗,对已产生抗-D的孕妇,应密切监测其抗-D水平,并在孕期及新生儿出生后给予正确治疗。     

与丈夫血型不合O型和/或RhD(-)孕妇产前血型免疫学抗体与HDN的关系

目的探讨与丈夫血型不合孕妇血型免疫学抗体效价与新生儿溶血病(HDN)发生的关系。方法对2709例O型Rh(+)孕妇血清IgG抗-A(B)和16例RhD(-)孕妇血清IgG抗-D效价进行检测与分析,并与临床资料进行比较研究。结果2709例O型孕妇血清中IgG抗-A(B)效价≥64者419例,占15.4%;16例RhD(-)孕妇血清中IgG抗-D效价≥64者16例,占100%。结论O型和/或RhD(-)孕妇与丈夫血型不合时,产前应及时检测血清中IgG抗体的水平;孕妇血清中IgG抗体效价的高低与HDN的发生成正比;HDN发生率与孕妇的年龄有关联;RhD(-)孕妇发生HDN与妊娠史有极大的因果关系,当抗体效价高时要及时用药治疗,降低其免疫性抗体效价,预防HDN的发生。     

多克隆抗RhD抗体的纯化和鉴定

摘要：目的：纯化人血清中多克隆抗RhD抗体,并对其进行鉴定。 方法：收集5例RhD阴性者血清,经不规则抗体筛查和确认后,按等比例混合,分别用RhD(-)A型红细胞和RhD(-)B型红细胞对血清中的抗A、抗B抗体于4 ℃过夜吸收处理;用RhD(+)O型红细胞制备血影细胞,以血影细胞作为抗原,吸附血清中抗RhD抗体,再用低pH值缓冲液洗脱、分离纯化抗RhD抗体,用DEAE-Sephadex A-50层析法提取其中IgG组分;分别用western blot、微柱法鉴定。 结果：5名研究对象血清中均含抗RhD抗体;血清中的抗A和抗B血型抗体被完全吸收,纯化后抗RhD抗体IgG组分效价达到1∶32。 结论：本实验获得了高效价的抗RhD多克隆抗体,为进一步利用噬菌体肽库获得RhD血型抗原表位研究等奠定基础。     

与丈夫血型不合O型和RhD阴性孕妇产前IgG抗体检测结果分析

目的探讨与丈夫血型不合O型和RhD阴性孕妇产前免疫性IgG抗体效价异常的意义。方法与丈夫血型不合O型孕妇2 027名和RhD阴性孕妇11名,采用微柱凝胶法分别于产前常规检测血清IgG抗-A(B)及抗-D效价。结果 2 027名O型孕妇产前血清IgG抗-A(B)效价≥64者1 076例,11名RhD阴性孕妇产前血清IgG抗-D效价≥16者8例。结论与丈夫血型不合O型和RhD阴性孕妇产前检测血清IgG抗-A(B)、抗-D效价,发现异常及时进行治疗,可预防新生儿溶血症的发生。     

RhD多肽抑制抗原抗体结合的效果

目的 评价RhD多肽抑制抗-D抗体与RhD抗原结合的效果。方法 根据随机噬菌体展示文库筛选得到RhD抗原模拟多肽的氨基酸序列,合成RhD多肽。利用生物信息学技术分析多肽理化性质及二级结构,微柱凝胶低离子抗人球蛋白卡凝集抑制试验验证有效多肽,显微镜观察多肽有效抑制浓度,流式细胞仪法检测多肽抑制效果。结果 Peptide 4噬菌体多肽最为稳定,未形成无规则卷曲与抗原性,且抑制IgG型单克隆抗-D与RhD抗原的反应效果较为明显,显微镜观察呈一定的剂量效应。流式细胞仪法显示当Peptide 4浓度达2.5 mg/50μL时对IgG型单克隆抗-D抗体与O型RhD阳性红细胞的凝集具有抑制作用。结论 Peptide 4为有效多肽,对抑制抗-D与RhD抗原的凝集具有一定效果,对RhD蛋白结构与功能研究及抗原性改造,提高临床输血的安全性等具有重要意义。     

RhD同种免疫有效性研究

目的通过RhD阳性红细胞免疫RhD阴性志愿者,对中国人群RhD主动免疫有效性进行研究,为我国生产抗-D免疫球蛋白提供理论支持。方法对14例RhD阴性志愿者进行RhD阳性红细胞免疫。初次免疫之后每3个月进行加强免疫。免疫后采集血样,检测抗-D效价、血常规与肝肾功能。采用t检验分析数据。结果初次免疫后应答率为4/14(28.6%),第1次加强免疫后免疫应答率为8/14(57.1%)。RhD阴性志愿者平均产生抗-D时间为初次免疫后4个月,加强免疫后7 d抗-D效价可达到峰值。加强频次≤2次与≥3次平均效价分别为16,1 024(P0.05)。结论本研究可在保障志愿者安全的前提下,为我国生产抗-D免疫球蛋白提供可靠的理论支持。     

流式细胞术产前非侵入性定量检测胎-母出血的研究

目的　建立产前非侵入性定量检测RhD阴性孕妇胎 母出血的流式细胞术实验方法。方法　RhD阴性红细胞与RhD阳性红细胞按一定比例混合 ,用抗 D作为一抗结合D+ 红细胞 ,以羊抗人IgGF(ab′) 2 FITC作为二抗 ,连接抗 D ,应用流式细胞术检测D+ 红细胞占D-红细胞的比例 ,建立抗 D及抗人IgGF(ab′) 2 FITC的量效关系 ,确定两者最佳使用剂量。在此实验条件下 ,进行流式细胞术检测值与实际值的相关性分析 ,判定可以检测出的D-红细胞中混有D+ 红细胞的准确出血量范围 ,应用建立的流式细胞术方法测定 9例RhD阴性孕妇外周血中的胎 母出血量。结果　抗 D的有效使用剂量为 1∶4 0 0稀释 ,10 0 μl/5× 10 6细胞 ;羊抗人IgGF(ab′) 2 FITC的有效使用剂量为 2 5 μg/5× 10 6细胞 ;流式细胞术测定比例与已知实际比例的相关系数r =0 998;准确检测的胎 母出血量为 (0 .6～ 30 )ml。 9例RhD阴性孕妇外周血中除 2例未检出D+ 细胞而无法判断外 ,其余 7例均检测出胎 母出血 ,胎 母出血量为 (1.2～ 6 .4 8)ml。结论　针对孕期发生胎 母出血反应的RhD阴性孕妇 ,可以应用流式细胞术方法在产前准确无损伤地定量检测胎 母出血量 ,指导临床预防、治疗Rh新生儿溶血病     

输血相容性试验室内质控品的研制

目的研制出简易可行,可在一般实验室使用的质控品。方法选择10人份健康献血者的浓缩红细胞(O/AB型RhD阳性)制备红细胞质控品,取新鲜血浆(O/AB型RhD阳性)和抗-D(IgG)人血清配制血浆质控品。对制备好的质控品进行性能评价,包括重复性、稳定性、最佳条件与常规条件下测定比较以及微柱凝胶法与凝聚胺法测定比较。结果质控品的批内重复性CV均10%,批间重复性差异无统计学意义(P0.05)。在不同保存时间,条件及方法的条件下,质控品检测结果均无明显变化(P0.05)。统计分析6个月的室内质控结果,在控率为100%。结论成功研制出输血相容性试验室内质控品,该质控品具有较好的重复性,稳定性及实用性。     

Quantification of IgG anti-D bound to D-positive red cells infused into D-negative subjects after intramuscular injection of monoclonal anti-D

SUMMARY. A flow-cytometric method was developed to determine the number of molecules of IgG bound to D-positive red blood cells (RBC) when sensitized with low plasma concentrations of IgG anti-D in the presence of an excess of D-negative RBC. D-positive RBC were infused into 12 D-negative male volunteers 2 days after intramuscular injection of monoclonal anti-D (BRAD-3, IgG3 or BRAD-5, IgG1). Blood samples were taken immediately before, 3 min and 3 h after injection of the RBC, incubated for 1 h at 37°C, washed, then incubated sequentially with FITC-conjugated anti-IgG, IgM monoclonal anti-D, biotin-conjugated anti-IgM, and R-phycoerythrin-conjugated streptavidin. To prepare a standard curve, O R₁R₂ RBC were incubated in duplicate with varying dilutions of BRAD-3 or BRAD-5, and RBC from one set were mixed with an excess of D-negative RBC (1:100) and labelled as above, while cell-bound IgG on the other set was quantified by ELISA. The test samples and standards were analysed by flow cytometry and the mean channel (FITC) fluorescence was plotted against molecules IgG/cell, from which the sensitization level of D-positive RBC in the test samples was determined. The use of IgM anti-D enhanced the discrimination between D-positive and D-negative RBC, especially when fewer than about 3000 molecules IgG/cell were bound. The assay was sensitive to about 1000 molecules IgG/cell. The sensitization levels of the D-positive RBC in samples taken 3 h after injection were found to be in the range 1500-10,000 molecules IgG anti-D per cell.     

Rh(D)阴性孕妇血液中胎儿微量Rh(D)红细胞与新生儿溶血病发病率的相关性研究

目的运用流式细胞术产前非侵入性检测Rh(D)血型不合导致新生儿溶血病,探讨该方法在临床的实用性。方法运用已建立的间接标记法(一抗为IgG抗-D,二抗为FITC-抗人IgG(γ),F(ab’)2)染色,采用流式细胞术检测Rh(D)阴性孕妇血液中胎儿微量红细胞。结果流式细胞术与血清学方法检测RhD血型不合导致新生儿溶血病差异不显著。追踪检测Rh(D)阴性孕妇体内胎儿微量Rh(D)红细胞所占比例数据显示,较早进行产前诊断能在一定程度上帮助预防、诊断HDN。结论建立了流式细胞术产前非侵入性检测Rh(D)血型不合导致新生儿溶血病的方法;孕妇在产前常规作流式细胞术检测,为Rh(D)血型不合导致新生儿溶血病提供了一种可靠的诊断依据。     

Secondary alloanti-D immunization post transfusion of “Asia type” DEL red blood cells

Background“Asia type” DEL red blood cells (RBCs) express a very weak D antigen and cannot be detected by routine RhD typing. Thus, it is routinely typed as D-negative (D–) blood group and transfused to D– recipients. Here we described a case of secondary alloanti-D immunization that was associated with transfusion of DEL RBCs to D– recipients and was initially considered as primary alloanti-D immunization.Case presentationA 44-year-old D– woman (G2P2) with adenomyosis and anemia underwent transabdominal hysterectomy. She received four units of D– RBCs before operation. Before transfusion, the alloantibody screening test was negative. Four days after the first transfusion, she needed another RBC transfusion. Unexpectedly, the routine pre-transfusion alloantibody screening test became positive and anti-D (titer, 128-fold) was identified, indicating an alloanti-D immunization. The anti-D developed four days after the first transfusion was unexplained, so alloantibody identification was performed on the sample collected before the first transfusion, and weak anti-D combined with anti-E, which was not detectable during the previous routine pre-transfusion alloantibody screening test with non-enzyme-treated screening cells, was identified using bromelain-treated panel cells. The remaining blood samples of first transfusion in bag tails from two donors were collected for RHD genotyping analysis. One donor was later identified as “Asia type” DEL having RHD* 1227 A/01 N.01 genotype.ConclusionCaution should be applied when we conclude that transfusion of “Asia type” DEL RBCs to true D– recipients could induce primary alloanti-D immunization, especially if the short time interval between transfusion and detection of anti-D is observed.     

Ultrastructural mapping of methyldopa and anti-D IgG erythrocyte antigen receptors.

The ultrastructural distribution pattern and site density of alpha-methyldopa immunoglobin G (alpha-MD IgG) on the red cell membrane was observed and compared with that of anti-D IgG, with ferritin-conjugated rabbit anti-human IgG and [125I]anti-D. alpha-MD IgG binds to all common types of human red cells, both Rho (D) positive and negative, to give a random, aperiodic distribution pattern grossly indistinguishable from the red cell D receptor site pattern. alpha-MD IgG inhibits the binding of [125I]anti-D to D-positive red cells when the reaction is controlled with respect to total reaction volume, ionic strength, and the appropriate concentrations of the two IgG reactants. To determine if a alpha-MD IgG binds to the D-antigen receptor, D-positive red cells were sensitized with alpha-MD and [125I]anti-D IgG spearately and with both IgG preparations. The cell-bound radioactivity served to identify what proportion of the total ferritin-labeled IgG sites were due to anti-D. With nonsaturating concentrations of anti-D the number of IgG sites observed was equal to the sum of the sites found when the red cell was sensitized separately with alpha-MD and anti-D IgG. With saturating concentrations of anti-D there was a reduction in the expected number of IgG sites, indicating that alpha-MD IgG was excluded from binding. There was no comparable interaction of alpha-MD IgG and anti-D IgG when D-negative red cells were used. The results obtained indicate that alpha-MD IgG does not bind to the D antigen. The interaction between alpha-MD IgG and anti-D IgG for binding sites on the red cell membrane may be due to the close physical proximity of the two receptors, so as to produce steric hindrance in binding of the two IgG preparations when both are present. The alpha-MD IgG receptor appears to be a part of the Rh antigen complex that occurs in both D-positive and D-negative red cells and probably contains receptors for other types of warm-antibody immune hemolytic anemias.     

全自动微柱玻璃珠技术提高血型相容性检测效能

探讨基于微柱玻瑞珠技术的ORTHO AutoVue Innova全自动血型及配血分析系统能否满足输血科血型血清学实验要求.用ORTHO AutoVue Innova全自动血型及配血分析系统和盐水介质法分别检测IgM类抗C、抗c、抗D、抗E和抗e系列稀释液各16份与相应抗原阳性的红细胞凝集反应的强度;用ORTHO AutoVue Innova全自动血型及配血分析系统、凝聚胺法和抗人球蛋白法分别检测IgG类抗D稀释液各16份与RhD阳性红细胞凝集反应的强度;再比较分析微柱玻璃珠法与传统对照实验的检测灵敏度.结果表明,ORTHO AutoVue Innova全自动血型及配血法对IgM类抗C、抗c、抗D、抗E和抗e的检测灵敏度分别为1:69.8、1:33.4、1:1448.1、1:139.6和1:32.0;盐水介质法的相应灵敏度分别为1:16.7、1:16.6、1:430.5、1:34.9和1:9.9.2种方法检测灵敏度的差异均具有统计学意义(IgM类抗C、杭c、抗D、抗E及抗e的t值分别为14.38、5.48、10.25、12.65和9.59,p均＜0.05).对于IgG类抗D微柱玻璃珠法、凝聚胺法和抗人球蛋白法的检测灵敏度分别为1:980.6、1:181.0和1:304.4,三种检测方法间灵敏度的差异具有统计学意义(F=51.15,p<0.01).结论:ORTHO AutoVue Innova全自动血型及配血法比传统方法具有更高的检测灵敏度,将其用于常规血型血清学检测是安全可靠的.     

Alloimmunization to D antigen and HLA in D-negative immunosuppressed oncology patients

D-negative patients may be divided into responders and nonresponders when immunized with D-positive red cells (RBC). Forty-nine D-negative oncology patients who received D-positive RBCs via platelet and white cell transfusions were studied to determine if nonresponders to D were likely to form lymphocytotoxic antibody (LCA). Nine patients developed anti-D in 16 to 390 days (mean = 112) after 2.6 to 481 ml (mean = 106) of D-positive RBCs. Forty patients had no evidence of anti-D after 0.8 to 535 ml (mean = 98) of D-positive RBCs and were followed for 14 to 1275 days (mean = 192). The anti-D group had no prior D-positive RBC transfusions, and two of five women making anti-D had previous pregnancies but no record of anti-D. LCA was found in four of nine (44%) patients with anti-D and in 12 of 40 (30%) patients without anti-D (p less than 0.50). Since both D and antigens HLA are considered highly immunogenic, it is of interest that the ability to form anti-D or LCA does not correlate. In fact, more patients (16/49; 32%) made LCA than anti-D (9/49; 18%). Of the 21 alloimmunized patients, 4 made both antibodies, while 17 had selective alloimmunization. It would thus appear that alloimmunization to D and HLA are not strongly linked and may indeed be unrelated.     

IgG1 and IgG3 anti-D in maternal serum and on the RBCs of infants suffering from HDN: relationship with the severity of the disease

BACKGROUND: Anti-D IgG antibodies that are responsible for severe cases of HDN belong chiefly to IgG1 and IgG3 subclasses. The relationship between the concentrations of IgG1 anti-D and IgG3 anti-D in maternal serum and the amount bound to the surface of infants' RBCs is not known. In addition, the contribution of the two subclasses to the severity of HDN is not well established. STUDY DESIGN AND METHODS: Blood samples from 40 infants suffering from severe forms of HDN due to anti-D were collected before transfusion together with sera from their respective mother. The amount of total anti-D IgG as well as IgG1 anti-D and IgG3 anti-D on infants' RBCs and the concentration in maternal sera were determined by ELISA. RESULTS: The median percentages of IgG1 anti-D and of IgG3 anti-D in maternal sera were 90 and 10 percent, respectively, whereas on infants' RBCs they were 97 and 3 percent, respectively. The differences between maternal and infantile percentages were significant (p < 0.001). IgG1 and IgG3 anti-D bound to infants' RBCs increased concomitantly with the concentration of IgG1 and IgG3 anti-D in maternal sera. The severity of HDN correlated positively with the concentration of IgG1 anti-D in maternal sera, but negatively with the amount of IgG3 anti-D bound to infants' RBCs. In addition, the existence of a high proportion of IgG3 anti-D in maternal serum was associated with a delayed risk of fetal anemia. CONCLUSION: The proportion of IgG3 anti-D relative to the total anti-D IgG on infants' RBCs is only one- third of the proportion present in maternal serum. The study of the correlations between the amount of IgG1 anti-D and IgG3 anti-D and the severity of HDN suggests that IgG1 anti-D are more important than IgG3 anti-D in the pathogenesis of fetal anemia.     

Immunospecific red cell binding of iodine 125-labeled immunoglobulin G erythrocyte autoantibodies

The primary interaction of autoantibodies with red cells has been studied by using labeled autoantibodies. Immunoglobulin G red cell autoantibodies obtained from IgG antiglobulin-positive normal blood donors were labeled with radioactive iodine and compared with alloanti-D with respect to their properties and binding behavior. Iodine 125-labeled IgG autoantibody migrated as a single homogeneous peak with the same relative mobility as human IgG on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric focusing pattern of labeled autoantibodies varied from donor to donor but was similar to that of alloanti-D, consisting of multiple IgG populations with isoelectric points in the neutral to alkaline range. 125I-autoantibody bound to all human red cells of common Rh phenotypes. Evidence for immunospecific antibody binding of the labeled autoantibody was based on variation in equilibrium binding to nonhuman and human red cells of common and rare phenotypes, enhanced binding after red cell protease modification, antiglobulin reactivity of cell-bound IgG comparable to that of cell-bound anti-D, and saturation binding in autoantibody excess. Scatchard analysis of two 125I-autoantibody preparations yielded site numbers of 41,500 and 53,300 with equilibrium constants of 3.7 and 2.1 X 10(8) L X mol-1. Dog, rabbit, rhesus monkey, and baboon red cells were antigen(s) negative by quantitative adsorption studies adsorbing less than 3% of the labeled autoantibody. Reduced ability of rare human D--red blood cells to adsorb the autoantibody and identification of donor autoantibodies that bind to Rh null red blood cells indicated that eluates contained multiple antibody populations of complex specificities in contrast to anti-D, which consists of a monospecific antibody population. Another difference is that less than 70% of the autoantibody IgG was adsorbed by maximum binding red blood cells as compared with greater than 85% for alloanti-D. Evidence obtained indicates that the red blood cell nonadsorbable IgG, some 30%, consists of other IgG populations, one of which represents IgG autoanti-idiotypes to the autoantibody.     

红细胞血型系统IgG型不规则抗体固相凝集法检测试剂盒的研发

目的研发基于固相凝集技术的红细胞血型系统IgG型不规则抗体检测试剂盒并评估其性能。方法采用单克隆抗人-RBC抗体包被96孔微板,按100μL/孔加入红细胞悬液形成细胞单层,经过裂解后形成红细胞膜抗原分子单层,加入保护液冷冻干燥后制备成检测用的反应微板;用梯度倍比稀释的多克隆羊抗球蛋白试剂(IgG+C3d/4)与IgG抗-D致敏的O型红细胞反应,筛选构建最佳指示系统;用不同批次的试剂盒检测低效价的IgG抗-D和抗-E,评估试剂盒在保存期内的抗原稳定性;与微柱凝胶抗球蛋白卡、进口同类试剂盒检测不同效价的抗体以测试3者的灵敏度;与进口同类试剂盒对350例血样标本进行检测,以明确2者的检测效能是否一致。结果 20μg/mL的抗人-RBC抗体溶液能够很好的固定红细胞膜抗原分子单层;稀释16倍的多克隆羊抗球蛋白试剂与致敏红细胞构建的指示系统指示效果最佳;试剂盒中冻干的红细胞膜抗原在6个月的保存期内保持稳定;本研究的试剂盒检测灵敏度高于Capture-R Ready Screen试剂盒和抗球蛋白卡;试剂盒与Capture-R Ready Screen对血样标本检测的阳性一致性百分率为98.0%,阴性一致性百分率为99.66%,总一致性百分率为99.43%。结论成功研发了基于固相凝集技术的用于红细胞IgG型同种免疫性不规则抗体检测的试剂盒,该试剂盒具有灵敏度好、保存期长、性能稳定等优势,与进口同类试剂盒等效。     

血液抗-D抗体检测在RhD阴性患者输注RhD阳性同型红细胞中的效果分析

目的研究RhD阴性患者输注与其血型相同的RhD阳性红细胞血液后,患者抗-D抗体的水平。方法收集该院2010年1月至2016年1月输注RhD阳性同型红细胞的20例RhD阴性患者的临床资料,分析其在输血前及输血后10、20、30、60、90d的抗-D抗体水平及RhD阳性患者的效价。结果输注RhD阳性同型红细胞血液的20例RhD阴性患者,在输血后90d内有6例患者表现为RhD阳性,其中男性RhD阳性率为25.0%(3/12)、女性RhD阳性率为37.5%(3/8)。3例RhD阳性的女性患者抗-D抗体效价分别为35、278、508。结论 RhD阴性患者输注RhD阳性同型红细胞血液,可刺激机体免疫机制,产生红细胞表面抗-D抗体,且部分RhD阴性患者的红细胞表型会发生改变。