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  • Immunohematology


Article | 15-February-2021

Heat elution: a modification of the Landsteiner-Miller method

between the antibody and its antigen is reversible and is affected by the ionic strength, temperature, and pH of the testing environment. The process of removing an antibody attached to red blood cells (RBCs), either in vivo or in vitro, is termed elution, and the recovered, concentrated antibody in solution is called an eluate. Heat elution, the first RBC elution procedure,2 was described by Landsteiner and Miller3 in their studies on chimpanzees. The original method involved a 5-minute incubation at

C. Dean-El, N. Quraishy

Immunohematology, Volume 35 , ISSUE 2, 45–47

Review | 01-December-2019

Cold acid elution (ELU Kit II)

Elution is a procedure for recovery of antibody attached to intact, immunoglobulin-coated red blood cells (RBCs) by disrupting the antigen–antibody bonds. The recovered antibody is collected in an inert diluent and is referred to as an eluate. Testing of an eluate may be desired to identify antibody(ies) coating the RBCs of patients with a positive direct antiglobulin test. Many types of elution procedures have been developed and described; however, an acid elution is suitable for

Monica Hinrichs, Monica A. Keith

Immunohematology, Volume 30 , ISSUE 3, 113–116

Article | 14-December-2020

Use of a modified acid/EDTA elution technique

A modification of the acid/EDTA elution technique was recently developed at the International Blood Group Reference Laboratory, Bristol, UK. By altering the volumes of reagents used, maximization of elution of antibody could be achieved without loss of red cell integrity.

Peter C. Byrne

Immunohematology, Volume 7 , ISSUE 2, 46–47

Article | 17-February-2021

Prevalence of DEL phenotype in D– blood donors in India

two homologous RH genes: RHD and RHCE. The high degree of homology and opposite orientation of the two genes leads to the production of numerous Rh variants.2 D variants are usually classified as weak D, partial D, and DEL. Unlike weak D and partial D, DEL represents a weakened form of D that cannot be detected by conventional serology and requires the use of an adsorption-elution method; therefore, it is DEL (D-elute).3,4 The first examples of DEL+ RBCs were reported in the early 1980s when blood

R. Chaudhary, S. Verma, A. Verma

Immunohematology, Volume 36 , ISSUE 4, 133–136

Article | 14-October-2020

The investigation of the significance of a positive direct antiglobulin test in blood donors

Sixty-two samples from 62 donors were investigated to determine the significance of warm IgG autoantibodies that were detected using a gel system during compatibility testing. The presence of autoantibodies on the red cells was confirmed by elution studies. Twelve of 23 strongly positive samples, 7 of 19 moderately positive samples, and 6 of 11 weakly positive samples were studied. The remaining nine samples were found positive during crossmatching, then negative when it was repeated.These nine

Marianna Bellia, John Georgopoulos, Vasilis Tsevrenis, Efrosini Nomikou, Niki Vgontza, I. Kontogpoulous-Griva

Immunohematology, Volume 18 , ISSUE 3, 78–81

Case report | 01-December-2019

Performance of an automated solid-phase  red cell adherence system compared with  that of a manual gel microcolumn assay for  the identification of antibodies eluted from  red blood cells

IgG antibodies coating red blood cells (RBCs) can be removed by elution procedures and their specificity determined by antibody identification studies. Although such testing is traditionally performed using the tube agglutination assay, prior studies have shown that the gel microcolumn (GMC) assay may also be used with comparable results. The purpose of this study was to compare an automated solid-phase red cell adherence (SPRCA) system with a GMC assay for the detection of antibodies eluted

Rachel H. Finck, Rebecca J. Davis, Shih-Mao Teng, Dennis Goldfinger, Alyssa F. Ziman, Qun Lu, Shan Yuan

Immunohematology, Volume 27 , ISSUE 1, 1–5

Article | 10-November-2020

Evidence that the low-incidence red cell antigens R1a and Lsa are identical

Testing of Ls(a+) and R1(a+) red cells with numerous antisera containing antibodies to low-incidence antigens indicated that these antigens are identical. This conclusion was confirmed by adsorption and elution tests, and supported by immunoblotting of Ls(a+) and R1(a+) cells with antibodies to giycophorin C and glycophorin D.

Leif Kornstad, Carole Green, Pertti Sistonen, Geoff Daniels

Immunohematology, Volume 12 , ISSUE 1, 8–10

case-report | 25-June-2021

B subgroup detection in a small hospital transfusion service

result between RBC and serum grouping tests. Subgroups of B are difficult to classify and include B3, Bx/Bweak (previously known as Bx), Bweak (previously known as Bm), and Bel phenotypes, according to the International Society of Blood Transfusion (ISBT).4,5 The B subgroups can be classified by their agglutination strength with anti-B, anti-A,B, and anti-H; the presence or absence of anti-B in the serum; the presence of B substance in saliva; the results of adsorption-elution testing; and molecular

E. Elardo, N. Elbadri, C. Sanchez, V. Powell, M. Smaris, Y. Li, J. Jacobson, T. Hilbert, T. Hamilton, D.W. Wu

Immunohematology, Volume 37 , ISSUE 2, 89–94

Case report | 27-December-2020

A case report: unusual Gerbich antibody in a patient with sickle cell anemia

A patient whose red blood cells (RBCs) typed as Ge:2,3 produced an alloantibody to a high-frequency antigen in the Gerbich system. This antibody was shown to be nonreactive with Ge: -2, -3 RBCs using adsorption-elution studies. A monocyte monolayer assay (MMA) suggested that transfusion of Ge:2,3 RBCs to this patient would have reduced in vivo survival.

Michael I. Gorman, Bobbye Woody

Immunohematology, Volume 5 , ISSUE 2, 55–57

Article | 20-December-2020

Du phenotyping by the rosette technique when the direct antiglobulin test is positive

). The microscopic “starry-sky” pattern typical for FMH-positive rosettes is easily differentiated from the uniform agglutination produced by the Du phenotype. The rosette technique was performed on 202 Rh-negative cord RBCs out of 4815 cord cells with a positive DAT. Parallel testing for Du by the chloroquine elution and antihuman globulin (AHG) test was done on 67 of the 202 cases. Four Du RBCs that were identified by the rosette technique were confirmed by the chloroquine Du technique

Jochewed Werch, Carolyn Todd, Marilyn K. Moulds

Immunohematology, Volume 6 , ISSUE 2, 44–46

Case report | 26-October-2019

Blocked D phenomenon and relevance of maternal  serologic testing

elution and gentle heat elution (at 56°C) confirmed the presence of anti-D on neonatal RBCs. The baby received two exchange transfusions with group O, D–, packed RBCs compatible with his own serum. Later, on day 3, the neonate’s mother was typed as group AB, D–, and her serum revealed the presence of alloanti-D, -C, and -S reactive in the anti-human globulin phase. The anti-D titer was 1024. This report highlights the “blocking” phenomenon caused by maternal anti-D in

Ashish Jain, Vijay Kumawat, Neelam Marwaha

Immunohematology, Volume 31 , ISSUE 3, 116–118

Article | 06-December-2020

Expression of B and H antigens on red cells from a group Bweak individual studied by serologic and scanning electron microscopic techniques

The proposita was classified as Bel, By, or Bm, Le(b+) by routine blood grouping and by adsorption/elution studies using anti-A and -B hyperimmune pregnancy sera. Red cells from the proposita adsorbed as much anti-B from the hyperimmune sera as did red cells from normal B individuals, but adsorbed less anti-A,B (group O serum). Saliva contained H, but not B, soluble substance. Red cells from the proposita and a normal B donor were sensitized with monoclonal A and B blood group antibodies

Hans Erik Heier, Leif Kornstad, Ellen Namork, Peder Østgard, Randi Sandin

Immunohematology, Volume 8 , ISSUE 4, 94–99

Article | 31-December-2020

Unreliability of Five Elution Techniques in an Attempt to Elute Autoanti-LW and Alloanti-LW

Rosia Nesbitt

Immunohematology, Volume 3 , ISSUE 3, 35–36

Article | 31-December-2020

An Example of Mild Hemolytic Disease of the Newborn Caused by Anti-Cob

A woman whose serum contained multiple alloantibodies delivered a full-term infant with mild hemolytic disease of the newborn. The direct antiglobulin test performed on the cord cells was positive with monospecific anti-IgG. An acid elution performed on the cord cells yielded anti-Cob. These findings were consistent with the presence of anti-Cob in the maternal serum. Neonatal clinical findings showed a mildly affected infant who demonstrated a moderate rise in total bilirubin and slight

Nancy B. Steffey, Mary A. Lieb

Immunohematology, Volume 3 , ISSUE 1, 9–10

Article | 03-November-2020

Implementation of gel column technology, including comparative testing of Ortho ID-MTS with standard polyethylene glycol tube tests

antibody enhancement. Tests were performed as described in the manufacturer’s guidelines and the current edition of the Technical Manual of the American Association of Blood Banks. Testing included antibody detection, antibody identification, direct antiglobulin tests (DATs), antigen phenotyping (K, Fya, Fyb, S, and s), and elution studies. These procedures were evaluated for sensitivity, specificity, and efficiency. Sixty-six samples that had been tested for antibody activity by PEG tube

Diane A. Derr, Stacy J. Dickerson, E.Ann Steiner

Immunohematology, Volume 14 , ISSUE 2, 72–74

Article | 14-October-2020

Anti-Mta associated with three cases of hemolytic disease of the newborn

showed that the mother’s serum contained anti-Mta . The father and all three children phenotyped as Mta+, while the mother was Mta–. Adsorption and elution experiments gave results which suggested that anti-Mta may be implicated in recurrent HDN in this family.

Carol C. Cheung, Daniel Challis, George Fisher, Susan J. Russell, Andrew Davis, Hayley Bruce, Julie Watt, Beng H. Chong

Immunohematology, Volume 18 , ISSUE 2, 37–39

Article | 16-October-2019

Separation of multiple antibodies by adsorption with allogeneic red blood cells

. Routinely, antibody identification practices in a blood bank comprise the testing of a patient’s plasma against reagent RBCs using standard agglutination and indirect antiglobulin methods.1 There are, however, instances when identification of multiple antibodies may be complicated and require additional serologic methods. When the presence of multiple antibodies is suspected, several methods—including neutralization of patient’s plasma, titration, elution, chemical or enzyme treatment of reagent RBCs

E.M. Ekema

Immunohematology, Volume 33 , ISSUE 4, 155–158

Case report | 09-October-2019

Autoanti-C in a patient with primary sclerosing cholangitis and autoimmune hemolytic anemia: a rare presentation

-reactive AIHA. Further testing showed the possibility of anti-C. The patient’s Rh phenotype was C+D+E–c– e+. Further testing with select cells, serial alloadsorption, and an elution confirmed anti-C specificity. The patient was transfused with two C–, crossmatch-compatible packed red blood cell units. The patient’s hemoglobin level and general condition showed improvement. This unique case report shows PSC associated with AIHA caused by autoanti-C. Usually, warm AIHA

Meenu Bajpai, Ashish Maheshwari, Shruti Gupta, Chhagan Bihari

Immunohematology, Volume 32 , ISSUE 3, 104–107

Article | 14-October-2020

Tube and column agglutination technology for autocontrol testing

agglutination tests. The tube method was carried out using low-ionic-strength solution (LISS). The direct antiglobulin test (DAT) was performed using the tube method, and further investigated with elution studies if warranted. Seventy-nine patient’s samples (7.74%) had a positive autocontrol: the gel test, 72 (91.13%); ReACT, 21 (26.58%); and the tube method, 27 (34.18%). Of the 79 positive autocontrols, 44 samples had a negative DAT. Of the samples with positive DAT results, only one possessed a

J.E. Courtney, J.L. Vincent, A.J. Indrikovs

Immunohematology, Volume 17 , ISSUE 2, 50–52

Article | 18-October-2020

Fyx is associated with two missense point mutations in its gene and can be detected by PCR–SSP

;) samples of Tanzanian origin were correctly typed and of 300 random donors of Caucasian origin with known Fy phenotype, only four out of 59 Fy(a+b–) donors showed the discrepant DNA-type Fy(a+b+). Serologic reinvestigation by adsorption and elution techniques confirmed weakly expressed Fyb antigen in these cases and DNA sequencing of the entire Duffy gene revealed identical point mutations in all of them. Specific PCR reactions were used to reinvestigate the C265T (Arg89Cys) and G298A (Ala100Thr

Christoph Gassner, Richard L. Kraus, Tadeja Dovc, Susanne Kilga-Nogler, Irene Utz, Thomas Mueller, Friedrich Schunter, Diether Schoenitzer

Immunohematology, Volume 16 , ISSUE 2, 61–67

Article | 20-December-2020

Human leukocyte antigens (HLA) class I (Bg) on red cells studied with monoclonal antibodies

immunoblotted with monoclonal antibodies, a complete 45 kDa intrinsic transmembrane heavy chain of HLA class I and a light chain of 11 kDa (ß2-M) were detected. Chloroquine treatment and acid elution of RBCs did not remove HLA class I but only ß2-M. As most antibodies recognize epitopes that depend on close association of dass I with ß2-M, the lost reactivity of treated RBCs may be understood.

Carolyn M. Giles

Immunohematology, Volume 6 , ISSUE 3, 53–58

Article | 26-October-2020

Naturally-occurring anti-Jka in infant twins

transfusions. Red blood cells (RBCs) from the patient and her sister typed as Jk(a-b+) by direct hemagglutination and this phenotype was confirmed by negative adsorption and elution studies. Both infants' plasma samples were strongly reactive with 20 examples of Jk(a+) RBCs and nonreactive with 20 examples of Jk(a-) RBCs by SPRCA assays. Anti-Jka was not detected in either twins' plasma by indirect antiglobulin tests by tube method in low-ionic-strength saline solution or polyethylene glycol, or

Dawn H. Rumsey, Sandra J. Nance, Mary Rubino, S. Gerald Sandler

Immunohematology, Volume 15 , ISSUE 4, 159–162

case-report | 25-June-2021

Neonatal testing leading to the identification of Bh (para-Bombay) phenotype in the mother: case report with review of the literature

substances in secretions.2 These individuals are homozygous for a nonfunctional FUT1 gene. The common mutations in FUT1 and FUT2 genes in a para-Bombay phenotype identified in recent years are summarized in Table 4. These individuals may express weak ABH substances on their RBCs by adsorbing a Type 1 precursor chain from the plasma. The negative results in the adsorption and elution in our case suggested that the RBCs were devoid of antigen expression. During the secretor status workup in this case, the

G. Mohan, A. Vaidya, S. Shastry

Immunohematology, Volume 37 , ISSUE 2, 59–63

Review | 14-March-2020

Laboratory methods for Rh immunoprophylaxis: a review

for detecting a fetomaternal hemorrhage in D+ mothers or when the D type of the fetus or newborn is D– or unknown. The acid-elution (Kleihauer- Betke) assay is a sensitive laboratory method for quantifying a fetomaternal hemorrhage, but it is tedious, often inaccurate, and difficult to reproduce. Flow cytometry, using anti-D or anti- hemoglobin F reagents, offers a more precise quantification of fetal RBCs in maternal blood. However, flow cytometry services for this function are available in

S. Gerald Sandler, Srividya Sathiyamoorthy

Immunohematology, Volume 26 , ISSUE 3, 92–103

Article | 03-November-2020

Autoimmune hemolytic anemia caused by warm-reacting IgM-class antibodies

confirmed as IgM by their ability to rebind to normal red blood cells (RBCs) after elution; the absence of small increases in RBC-bound IgG and IgA was shown by a sensitive enzyme-linked antiglobulin test. Patient 1 was a 64-year-old female with non-Hodgkin’s lymphoma, with a hemoglobin of 50 g/L and haptoglobin of < 0.1 g/L. Direct antiglobulin tests were positive for IgM, C3d, and C3c; only IgM was present in an eluate. The serum contained a weak autoantibody at 37°C and tests for

R.J. Sokol, D.J. Booker, R. Stamps, S. Sobolewski, A.P. Haynes

Immunohematology, Volume 14 , ISSUE 2, 53–58

Review | 09-October-2019

DEL phenotype

DEL red blood cells (RBCs) type as D– by routine serologic methods and are transfused routinely, without being identified as expressing a very weak D antigen, to D– recipients. DEL RBCs are detected only by adsorption and elution of anti-D or by molecular methods. Most DEL phenotypes have been reported in population studies conducted in East Asia, although DEL phenotypes have been detected also among Caucasian individuals. Approximately 98 percent of DEL phenotypes in East Asians

Dong Hyang Kwon, S. Gerald Sandler, Willy Albert Flegel

Immunohematology, Volume 33 , ISSUE 3, 125–132

Case report | 01-December-2019

Molecular analysis of patients with weak D and serologic analysis of those with anti-D (excluding type 1 and type 2)

included autologous controls, direct antiglobulin test, elution, and titration of anti-D before and after adsorption of serum onto autologous RBCs. From molecular analyses, 459 individuals exhibited a weak D type. We described seven novel RHD variant alleles. The most frequent types of weak D were type 1 (30.1%), type 2 (23.7%), type 4.0 (10.2%), type 4.2.2 (20.3%), type 11 (3.9%), and type 15 (3.7%). Anti-D was identified in the sera of 9 of 47 individuals with weak D type 4.0, in 14 of 93 with weak D

Bach-Nga Pham, Michèle Roussel, Dominique Gien, Maryline Ripaux, Carine Auxerre, Pierre-Yves Le Pennec, Christine Andre-Botte

Immunohematology, Volume 29 , ISSUE 2, 55–62

Report | 14-March-2020

Absence of hemolytic disease of fetus and newborn despite maternal high-titer IgG anti-Ku

elution of RBC antibodies, and antigen typing. A gravida 3, para 3 (G3P3) woman was first evaluated in 2006 and was found to have an IgG RBC antibody that reacted against all panel RBCs in the anti-human globulin phase. A panel of RBCs treated with DTT did not react with the antibody.  The antibody failed to react with one example of K0 RBCs. The patient’s RBCs typed negative for the following Kell blood group antigens: KEL1, KEL2, KEL3, KEL4, KEL6, KEL7, KEL11, KEL13, and KEL18. These

Ram M. Kakaiya, Angelica Whaley, Christine Howard-Menk, Jigna Rami, Mona Papari, Sally Campbell-Lee, Zbigniew Malecki

Immunohematology, Volume 26 , ISSUE 3, 119–122

Article | 15-April-2020

Efficacy of murine monoclonal antibodies in RBC phenotyping of DAT-positive samples

Determining the phenotype of patient RBCs that are positive by the DAT may prove problematic. Antigen typing of RBCs coated with IgG requires direct agglutinating reagents or chemical treatment (such as chloroquine diphosphate [CDP] or citric acid) to remove sufficient IgG to permit testing with IAT-reactive reagents. The citric acid elution method is commonly used in the United States; however,antigens in the Kell system are altered to the extent that they may appear to be absent by this

Edmond Lee, Kevin Hart, Gordon Burgess, Gregory R. Halverson, Marion E. Reid

Immunohematology, Volume 22 , ISSUE 4, 161–165

Case report | 01-December-2019

Serologic and molecular characterization of D variants in Brazilians: impact for typing and transfusion strategy

%), 13 weak D type 3 (7.8%), and 2 weak D type 5 (1.2%) alleles were found. Among the partial D samples, 49 type 4.0 weak partial D (36%), 9 DAR (6.6%), 24 DFR (17.6%), 6 DBT (4.4%), 1 DHMi (0.73%), 26 DVI (19%), 14 DVa (10.3%), 5 DIVb (3.7%), and 2 DVII (1.5%) were observed. Two samples identified as DEL by adsorption-elution were characterized by molecular analyses as RHD(IVS5–38DEL4) and one sample was characterized as RHD(K409K). One sample was characterized as DHAR, a CE variant positive

Débora Castilho Credidio, Jordão Pellegrino Jr., Lilian Castilho

Immunohematology, Volume 27 , ISSUE 1, 6–11

Article | 16-October-2019

Assessment of common red blood cell pretreatments to yield an accurate serologic antigen phenotype compared with genotype-predicted phenotype

others12 and is a common cause of Fyb typing discrepancies. This scenario may result in interpreting a patient with Fyx as Fy(b–) and may complicate provision of antigen-matched RBCs for transfusion-matching protocols requiring FY status. More recently, a similar variant was found that can cause weakening of Fya.13 Furthermore, flow cytometry of EGA-treated RBCs suggests that samples negative by tube DAT after EGA elution may still have trace amounts of RBC-bound IgG detectable only by flow cytometry

T. Horn, J. Hamilton, J. Kosanke, V.W. Hare, W. Kluver, W. Beres, S. Nance, M.A. Keller

Immunohematology, Volume 33 , ISSUE 4, 147–151

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