微柱凝胶法与试管法检测新生儿溶血病IgG抗体效价的比较

目的：探讨微柱凝胶法在新生儿溶血病实验诊断中测定孕妇血清中IgG抗体效价的可行性及方法学。方法：孕妇血清用2-Me处理做倍量稀释后，采用微柱凝胶法测定IgG抗体效价。同时与试管法进行对照比较。结果：用微柱凝胶法和试管法同时检测孕妇血清中的IgG抗体效价，2位检验者检测结果符合率试管抗人球蛋白法为79．3％，微柱凝胶法为95．1％，效价〈64时，微柱凝胶法灵敏度75．6％（62／82），试管抗人球蛋白法为86．6％（72／82），效价≥64时，微柱凝胶法灵敏度24．4％（20／82），试管抗人球蛋白法为13．4％（11／82）。采用Х^2。检验，Х^2=1.64，结果差异无统计学意义（P〉0．05）。结论：应用微柱凝胶法检测孕妇血清IgG抗体效价，能够使实验方法简单化、自动化，使实验操作标准化，判断结果直观，与试管法相比减少了肉眼判断结果时的人为误差。     

孕产妇血型抗体效价测定在产前诊断中的意义

目的：通过对孕产妇血清中血型抗体效价的测定,旨在了解异常IgG抗-A（B）或抗-D水平在孕妇中的分布情况,探讨孕妇血清中IgG类血型抗体效价与新生儿溶血病（HDN）之间的关系,为预防及治疗HDN提供重要的实验依据。方法：采用凝聚胺法分别对1316例0型血孕妇和64例Rh（D）阴性血孕妇血清中IgG抗-A（B）和抗-D水平做出测定。对新生儿运用微柱凝胶法做HDN溶血3项检测。结果：①在所检测的1316例0型血孕妇中,IgG抗-A测定有802例,效价≥64为129例,异常检出率占总抗-A的16．1％;IgG抗-B测定有744例,效价≥64为135例,异常检出率占总抗B的18．1％。其中89例IgG抗A（B）≥64O型血孕妇所生新生儿中,有22例患HDN,阳性率占24．7％;②64例Rh（D）阴性血受检孕妇中,5例检测出IgG抗-D抗体,抗体检出率占7．8％,新生儿中有3例患Rh—HDN。结论：①HDN的发病率随着母体内的IgG类血型抗体水平增高而增大;②对于Rh（D）阴性的孕妇,IgG抗-D效价与孕次成正相关。     

O型血浆和红细胞上清液抗体效价与时间消长性研究

目的：了解O型供血者全血血浆和悬浮红细胞上清液抗体效价与保存时间相关性。方法：采用盐水法检测血浆和红细胞上清液IgM抗A、抗B效价，用2-巯基乙醇（2-Me）破坏IgM抗体后，用抗人球蛋白法检测血浆和红细胞上清液IgG抗体效价。结果：在各保存的时间段中，O型全血血浆IgG抗A和kG抗B效价积分分别与悬浮红细胞清液IgG抗A和IgG抗B效价积分互相比较差异有统计学意义（P〈0.05～0．01）；同样，IgM抗A和IgM抗B效价积分分别与悬浮红细胞上清液IgM抗A和IgM抗B效价积分互相比较差异也有统计学意义（P〈0．05～0．01）。另外，O型全血血浆和悬浮红细胞上清液中IgM抗A和IgM抗B效价积分均明显高于IgG抗A和IgG抗B效价积分分别相互比较差异有统计学意义（P〈0．05～0．01）。全血血浆和悬浮红细胞上清液抗体效价随着保存期的延长至15d均呈阶梯式的下降，A、B组O型全血血浆和悬浮红细胞上清液中IgM抗A和抗B效价积分、IgG抗A和抗B效价积分分别与C、D、E、F组相互比较差异有统计学意义（P〈0．05～0．01）。结论：在紧急情况输血时，可酌情输注O型CPD保养液保存时间为15d以上的悬浮红细胞，可能不会引起不同红细胞血型受血者溶血性输血反应。     

微柱凝胶法检测孕妇IgG抗体与ABO-HDN发病率相关性

目的：母婴血型不合时,探讨微柱凝胶法检测O型孕妇产前IgG抗A（B）抗体水平与产后新生儿溶血病（HDN）的发病关系。方法：对有孕产史的ABO血型不合的O型孕妇168例,孕22～28周时取其血清用2-Me灭活处理后,测定其IgG抗A（B）抗体效价;分娩后取新生儿（脐）血液进行ABO血型鉴定,血清胆红素测定及新生儿3项溶血试验。对IgG抗体效价与HDN发生情况进行分析比较。结果：168例有孕产史的孕妇IgG抗A（B）抗体效价≤64者15例,128～256者47例,〉256者106例。其中新生儿溶血3项确诊ABO-HDN65例（38.7%）。新生儿ABO-HDN的发病率随孕妇体内IgG抗体效价的升高而升高。结论：微柱凝胶法对产前孕妇进行IgG抗体水平测定可以预测HDN的发生,对早期预防治疗HDN具有重要意义。     

微粒蛋白质组学研究进展

目的：比较高效价抗人球（效价≥1：512）蛋白、抗人球蛋白试剂（效价≥1：4）（低效价）与微柱凝胶法在自身免疫性溶血性贫血（AIHA）实验室检测中的优势。方法：首先以微柱凝胶法直抗试验筛选阳性血液标本,然后对上述标本分别用高效价抗人球蛋白（效价≥1：512）、抗人球蛋白试剂（效价≥1：4）做直抗半定量试验,γ中和试验。结果：微柱凝胶法易出现假阳性,抗人球蛋白试剂（效价≥1：4）易出现假阴性。高效价抗人球（效价≥1：512）未发现假阳性,而且凝集梯度明显,易于结果判断分析。结论：高效价抗人球（效价≥1：512）在AIHA直抗试验中,较抗人球蛋白试剂（效价≥1：4）与微柱凝胶法具有结果更加准确的明显优势。     

孕妇产前抗体效价预测新生儿溶血病

目的：探讨O型及Rh阴性孕妇血清中抗体效价来预测新生儿溶血病的关系。方法：采用血型血清学方法,对1256对夫妇血型不合的O型及Rh阴性孕妇检测ABO、RhD血型、抗体筛选、抗体效价。效价≥64的建议服中药治疗,定期检测抗体效价。对新生儿（出生0～7d）发生黄疸后,抽静脉血检测溶血3项。结果：1244名O型孕妇血清中IgG抗-A（B）效价≥64者564例,占45.3%;对12名Rh阴性孕妇血清中产生抗D抗体5例,效价均≥64,占41.6%,7例抗体筛选阴性。IgG抗-A（B）效价≥64并发生ABO-HDN占49.5%。ABO-HDN占产前的比例（279/1244）为22.4%。结论：动态监测夫妇血型免疫抗体效价,及时采取措施以降低孕妇体内IgG抗体效价,降低新生儿发病率,减少并发症,提高人口素质。     

微柱凝胶法检测Rh血型免疫性抗体的分析

目的：探讨Rh血型免疫性抗体筛查及特异性鉴定在临床输血和新生儿溶血病预防中的意义,观察微柱凝胶法检测Rh血型免疫性抗体的敏感性。方法：应用微柱凝胶法对患者输血前和孕妇产前做Rh血型免疫性抗体筛查,对筛查阳性者进一步做抗体特异性鉴定及效价测定。结果：在9878例患者中检出Rh血型免疫性抗体阳性86例（0.87%）,其中抗D12例（14.0%）,抗E25例（29.1%）,抗C11例（12.8%）,抗c9例（10.5%）,抗e4例（4.7%）,抗Ce7例（8.1%）,抗CE5例（5.8%）,抗CD6例（6.9%）,抗cD3例（3.5%）,抗cE4例（4.7%）。结论：在输血前和产前对拟输血患者和夫妇血型不合的孕妇进行Rh血型免疫性抗体筛查、特异性鉴定及效价测定,对确保输血安全及优生优育有重要的临床意义。     

流式细胞术检测低效价Rh抗体的研究

目的:建立流式细胞仪检测Rh血型低效价抗体的方法。方法:将不同效价IgG抗-D试剂与标准化人源Rh(D)阳性红细胞混合,加入异硫氰基荧光素标记的鼠抗人IgG二抗,用流式细胞仪检测样本抗-D抗体含量。检测结果与抗球蛋白法进行对比。结果:2种方法对于抗-D抗体水平变化趋势的检测结果一致,但流式细胞术可检测到128倍和256倍稀释的低效价抗体,其特异性、灵敏性优于抗球蛋白法。结论:流式细胞术可以用于低效价IgG抗-D抗体的检测,且能将其定量,效果优于传统方法。     

抗绿脓杆菌外毒素—A血浆（马）抗体效价检测方法的探讨

目的：为了探讨在生产条件下采用哪种方法来检测抗绿脓杆菌外毒素—A血浆（马）的抗体效价更为可行。方法：本文对小鼠中和试验，双向琼脂扩散试验及胶乳凝集试验进行了比较研究。结果实验显示，小鼠中和试验和双向琼脂扩散试验特异性良好，结果稳定，但前者灵敏度比后者更高。而胶乳凝集试验由于胶乳抗原不易保存，其结果稳定性较差。结论：我们认为小鼠中和试验适用于大规模生产时采用，双向琼脂扩散可作为初步效价的检测。     

抗-A、抗-B效价质控品的制备及初步应用

目的:制备抗-A、抗-B效价检测质控品并进行初步应用。方法:倍比稀释抗-D单克隆抗体,确定抗体最佳稀释度,实验组取质控品1.56μL加入200μL血清中,对照组取1.56μL NS加入200μL血清中,根据凝集强度判断实验是否在控,并比较2组抗体效价差异。取不同时间点冷藏保存的质控品,检测其IgM、IgG抗-D抗体效价,评估质控品性质是否稳定。结果:质控品IgM抗体效价为1∶512,最佳稀释度为1∶256,实验组、对照组抗-A、抗-B效价差异无统计学意义,不同时间点保存的质控品抗体效价差异无统计学意义。结论:自制抗体效价质控品具有良好稳定性与可靠性,可有效发现IgM抗体破坏不完全、稀释度误差、凝集度判断标准不同造成的效价检测失控问题。     

Comparison of conventional tube test with diamed gel microcolumn assay for anti-D titration

Anti-D titration is the first step in the evaluation of the RhD-sensitized patient. Traditionally, anti-D titration has been performed by tube agglutination. Gel microcolumn assay is a method that has gained widespread usage throughout the world, mainly for ABO/Rh typing, unexpected antibody screening and direct antiglobulin tests. As gel assay has become widely used as a routine method to detect red blood cell alloantibodies, a critical anti-D titer needs to be established. Seventy-nine known blood samples with anti-D (titers 1-32) were titrated simultaneously by the conventional tube test and the gel microcolumn assay. Red blood cells (R0r phenotype) were used, with a final concentration of 3% for tube and 0.8% for gel. Serial twofold dilutions (2-2.048) were prepared for each technique, followed by reading in antiglobulin phase. Anti-D titration in the gel microcolumn assay showed significantly higher titers (mean 3.4-fold) than the conventional tube test in all samples studied. Based on these data, it was not possible to determine a critical titer for anti-D titration by the gel microcolumn assay.     

Comparison of conventional tube test with diamed gel microcolumn assay for anti‐D titration

Summary Anti‐D titration is the first step in the evaluation of the RhD‐sensitized patient. Traditionally, anti‐D titration has been performed by tube agglutination. Gel microcolumn assay is a method that has gained widespread usage throughout the world, mainly for ABO/Rh typing, unexpected antibody screening and direct antiglobulin tests. As gel assay has become widely used as a routine method to detect red blood cell alloantibodies, a critical anti‐D titer needs to be established. Seventy‐nine known blood samples with anti‐D (titers 1–32) were titrated simultaneously by the conventional tube test and the gel microcolumn assay. Red blood cells (R₀r phenotype) were used, with a final concentration of 3% for tube and 0.8% for gel. Serial twofold dilutions (2–2.048) were prepared for each technique, followed by reading in antiglobulin phase. Anti‐D titration in the gel microcolumn assay showed significantly higher titers (mean 3.4‐fold) than the conventional tube test in all samples studied. Based on these data, it was not possible to determine a critical titer for anti‐D titration by the gel microcolumn assay.     

微柱凝胶法与试管法检测新生儿溶血病免疫性抗体的比较

目的：比较微柱凝胶法与试管法对母婴血型不合新生儿溶血病（HDN）免疫性抗体的检出率。方法：对临床表现为高胆红素血症、疑似HDN的患儿血标本同时用微柱凝胶法和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,并对患儿阳性血标本进行血型不规则抗体特异性鉴定及其效价测定。结果：在275例临床表现为高胆红素血症、疑似HDN的患儿血标本中,用试管法检出直接抗人球蛋白试验阳性180例（65.5%）,抗体游离试验阳性197例（71.6%）,抗体放散试验阳性210例（76.4%）;用微柱凝胶法检出直接抗人球蛋白试验阳性224例（81.5%）,抗体游离试验阳性238例（86.5%）,抗体放散试验阳性251例（91.3%）。微柱凝胶法比试管法的凝集强度高1＋～2＋。结论：微柱凝胶法的敏感性略高于试管法,具有操作简便,影响因素少,易于标准化,结果客观及保存时间长等优点。     

Comparative Evaluation of Five Different Methods of Anti‐ABO Antibody Titration: An Aid for ABO‐Incompatible Organ Transplants

The accurate estimation of ABO antibody titers is of the utmost importance in organ transplants involving ABO incompatibility. We aim to compare five different methods of titration and analyze the data. Samples of 48 O group blood donors who donated during the month of December 2015 to January 2016 in our institution were subjected to ABO antibody titration by five different methods: immediate spin (IS) tube titer, antihuman globulin phase tube titer, Coomb's gel card titer, gel card titer after dithiotreitol (DTT) treatment of plasma, and the solid phase red cell adherence method. The mean number of titer serial dilution steps in the different titer estimation methods was compared using the paired t‐test and McNemar test. A correlation between the methods was tested using Spearman's rho and kappa statistics. The median antiglobulin (AHG) phase tube titers were found to be the highest anti‐A (128) and anti‐B (192) titers. Significant differences in the ABO antibody titer readings among the five different methods were noted. Titers were reduced by DTT treatment in nearly 50% samples tested for both anti‐A and anti‐B titers. Average agreements between the DTT‐applied AHG phase gel card titers and the solid phase red cell adherence (SPRCA) titers was observed for anti‐A (κ = 0.473) and anti‐B (κ = 0.530). The AHG phase tube and gel cards titers showed poor agreements. There are differences in the interpretability of the ABO antibody titer among different techniques. Consistent and uniform application of the method for titration throughout the treatment of a patient is highly essential.     

应用微柱凝胶技术筛查新生儿ABO溶血病及结果分析

目的：探讨微柱凝胶技术在新生儿溶血病（HDN）筛查中的应用以及结果分析。方法：采用微柱凝胶技术对633例疑似HDN患儿进行检测,用经典抗球蛋白法做对照。结果：633例筛查实验微柱凝胶技术阳性率为30.5%,28%为ABO-HDN,2.5%为RH-HDN,其中直接抗球蛋白实验阳性率为6.8%,患儿红细胞抗体放散实验阳性率为30.5%（抗A抗体阳性89例,抗B抗体阳性45例,抗AB抗体阳性43例,抗D抗体阳性10例,抗E抗体阳性6例）,血清游离抗体鉴定阳性率为13%,而经典抗球蛋白法的直接抗球蛋白实验阳性率为3%,患儿红细胞抗体放散实验阳性率为仅为24.7%。结论：在温州地区的HDN以ABO-HDN最为常见,抗体分布特征为抗A抗体〉抗B抗体〉抗AB抗体〉抗D抗体〉抗E抗体。新生儿溶血微柱凝胶技术能快速准确检测出HDN,具有很高的灵敏度,并能够检出经典抗球蛋白法所不能检出的阴性标本,是直观可靠的方法。     

43例待产孕妇Rh血型抗体筛查阳性分析

目的：研究多次妊娠妇女体内Rh血型抗体与新生儿溶血病的关系。方法：运用微柱凝胶免疫法鉴定Rh抗体和分型,抗人球试验法测定抗体效价。结果：待产孕妇16845例,检出意外抗体48例,其中43例为Rh血型抗体且夫妇ABO顺式相容。分析抗体特异性种类和效价,抗D抗体8例,效价8～512;抗c抗体5例,效价16～128;抗E抗体16例,效价8～64;抗C抗体12例,效价4～8;抗e抗体2例,效价8。由此得出Rh血型抗原性强弱为D〉c〉E〉C〉e。结论：多次妊娠妇女体内Rh相关抗体效价强弱是引起新生儿溶血的原因之一。     

Application of the Gel Test in IgG Subclassing – A Comparison of Two Agglutination Assays

Background and objectives: IgG subclasses of anti-D seem to play some role in hemolytic disease of the newborn, but there has been disagreement as to its exact nature. The aim of our study was to evaluate a new technique for IgG subclassing of anti-D and to compare it to an established test. Materials and methods: In 31 cases of RhD immunization, we have compared two simple agglutinating assays for subclassing anti-D: an established V-well microtiter tray assay and a new gel test assay. Polyclonal rabbit anti-IgG agglutinating antibodies were tested against sensitized D-positive red blood cells by both assays. Results: The anti-D concentration in tested sera varied between 0.1 and 81 μg/ml (0.5–405 IU/ml). Both assays were simple to perform and the results correlated well. Conclusions: The gel test had two advantages: quick performance and easy interpretation.     

新生儿溶血病血清学检查2种方法的对比研究

目的：对比研究微柱凝胶法和抗人球蛋白试验在新生儿溶血病（HDN）血清学检查中的应用。方法：选择母婴ABO血型不合的新生儿血液标本80份。每份标本均用微柱凝胶法和抗人球蛋白试验进行HDN“三项检查”：直抗试验、游离试验和释放试验。并将数据进行统计学处理。结果：直抗试验中2种方法均无阳性结果；游离试验中2种方法比较差异无统计学意义（P〉0．05）；释放试验中2种方法比较差异有统计学意义（P〈0．05）。结论：HDN“三项检查”中微柱凝胶法优于试管法，快速、简便，标本量少，重复性强，结果判定直观，敏感性强。     

Evaluation of the Microcolumn Technology for Pretransfusion Testing in Multiple Myeloma Patients

Objective: Microcolumn tests are useful for serological investigations, although because of their high sensitivity, false-positive results might be expected, e.g. in hypergammaglobulinemia. The aim of this study was to evaluate these tests in multiple myeloma. Methods: Pretransfusion testing was done in 80 patients with multiple myeloma using microcolumn and traditional tube tests. Results: All sera were negative in microcolumn indirect antiglobulin test and enzyme test, positive in 58% of samples in the enzyme tube test. The microcolumn direct antiglobulin test was positive in about 40% of samples but never in the tube direct antiglobulin test. This was not due to the presence of autoantibodies but to nonspecific binding of immunoglobulins related to their concentration in sera. Conclusion: Microcolumn tests appeared to be useful for pretransfusion testing in multiple myeloma in spite of positive autocontrols.