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  • Immunohematology

 

Article | 15-February-2021

Albumin-indirect antiglobulin test

reagent RBC suspension. Centrifuge, and perform immediate spin reading (if desired). Add two drops albumin (per MI). Incubate 30–60 minutes at 37°C (per MI). Centrifuge, and perform 37°C reading. Wash three to four times in isotonic saline. Add one to two drops AHG reagent (per MI). Centrifuge, and perform AHG reading. Compiled from Fung et al.6 RBC = red blood cell; MI = manufacturer’s instructions; AHG = antihuman globulin. Indications The albumin-indirect antiglobulin test (IAT) is a

J.R. Hamilton

Immunohematology, Volume 35 , ISSUE 2, 63–64

Article | 16-February-2021

Saline–indirect antiglobulin test

Principle The saline–indirect antiglobulin test (IAT) or saline– indirect Coombs test refers to a test incubated at 37°C without enhancement media before washing and the addition of antihuman globulin (AHG) reagent. Saline-IAT tests rely solely on the binding kinetics between plasma antibodies and antigens expressed on red blood cells (RBCs) suspended in isotonic saline. In this environment, the Na+ in the saline may partially neutralize the oppositely charged RBC membrane, blocking the

J.R. Hamilton

Immunohematology, Volume 35 , ISSUE 4, 156–158

Article | 26-October-2020

EDTA/glycine-acid versus chloroquine diphosphate treatment for stripping Bg antigens from red blood cells

EDTA/glycine-acid (EGA) has been reported to remove IgG-bound antibodies from red blood cells (RBCs) and to denature Kell system and Era antigens. EGA-treated RBCs were tested in parallel with chloroquine diphosphate (CDP)-treated RBCs to evaluate whether EGA would remove Bg antigens from RBCs as efficiently as CDP. Fifty-seven serum/plasma samples containing known Bg antibodies were tested with untreated Bg+ RBCs, EGA-treated Bg+ RBCs, and CDP-treated Bg+ RBCs by an indirect antiglobulin test

Kayla D. Champagne, Peggy Spruell, Jane Chen, Leslie Voll, Gloria Schlanser

Immunohematology, Volume 15 , ISSUE 2, 66–68

Report | 29-October-2019

Indirect antiglobulin test-crossmatch using low-ionic-strength saline–albumin enhancement medium and reduced incubation time: effectiveness in the detection of most clinically significant antibodies and impact on blood utilization

Indirect antiglobulin test-crossmatch (IAT-XM) using enhancement media such as low-ionic-strength saline (LISS) and polyethylene glycol (PEG) usually requires 15 minutes of incubation. These methods are necessary when testing samples from blood recipients who have a higher risk of alloimmunization. In emergency situations, IAT-XM can be time-consuming and can influence presurgery routine, resulting in more red blood cell (RBC) units being tested and stored to avoid the transfusion of 

Carla Luana Dinardo, Sílvia Leão Bonifácio, Alfredo Mendrone Júnior

Immunohematology, Volume 30 , ISSUE 1, 1–5

Article | 16-October-2019

Dithiothreitol treatment of red blood cells

treatment of multiple myeloma, targets the CD38 antigen on malignant plasma cells. CD38 is also weakly expressed on all normal RBCs, including those in RBC reagents used in pretransfusion testing. This expression complicates the identification of clinically significant RBC antibodies because the plasma/serum of such patients will react with most or all RBCs in antibody detection and identification tests performed by the indirect antiglobulin test.5 Because CD38 configuration depends on disulfide bonds

C.B. Bub

Immunohematology, Volume 33 , ISSUE 4, 170–172

Article | 10-November-2020

The impact of a gel system on routine work in a general hospital blood bank

The gel system has been reported to be more sensitive and specific than the conventional tube indirect antiglobulin test (IAT) for anti­body screening. However, a major concern about the gel system is its cost. A cost analysis study was therefore conducted at our hospi­tal. The gel system costs more than the conventional tube IAT per test; however, the total staff and reagent costs per year were about equal, because of staff savings. Workload and cost per patient requir­ing blood

A. Chan, H.F. Wong, C.H. Chui, L. Wong, G. Cheng

Immunohematology, Volume 12 , ISSUE 1, 30–32

Article | 17-November-2020

The first example of anti-Gya detected in Hong Kong

The incidence of anti-Gya is known to be extremely rare, and only a few examples have been reported in Japanese persons and in Caucasians. This case history reports the first example of anti-Gya detected in a Chinese person. The propositus is an 83-year-old male patient whose anti-Gya was revealed because of incompatibility of all units of blood tested by a saline indirect antiglobulin test (SIAT). Although the antibody had a titer of 512 by SIAT, 10 units of Gy(a+) red cells were transfused

King Hang Mak, Derek S. Ford, Gregory Cheng, Che Kit Lin, Cindy Yuen

Immunohematology, Volume 11 , ISSUE 1, 20–21

Article | 26-October-2020

Precipitation of serum proteins by polyethylene glycol (PEG) in  pretransfusion testing

PEG was added. Rapid recognition of this phenomenon could prevent delay in the selection of blood for transfusion by substituting PEG-indirect antiglobulin test (IAT) with another technique such as low-ionic-strength solution and by increasing the number of washes prior to addition of the antiglobulin reagent.

Jack Hoffer, William P. Koslosky, Elizabeth S. Gloster, Therese M. Dimaio, Marion E. Reid

Immunohematology, Volume 15 , ISSUE 3, 105–107

Article | 14-October-2020

One thousand seventy antibodies detected only by a 2-stage papain test: wanted and unwanted positive reactions

Despite the wide use of the antibody detection test for unexpected antibodies, controversy still remains regarding the use of enzymetreated red blood cells. Over a 6-year period, 72,573 samples from 49,863 patients submitted for pretransfusion compatibility testing were examined for unexpected antibodies. The antibody detection tests included a low-ionic-strength solution (LISS) indirect antiglobulin test and a two-stage papain (2SP) test. One thousand and seventy of the 2267 (47%) antibodies

Carmen Martin-Vega, Dolores Castella, Joan Cid, Marta Panadés

Immunohematology, Volume 17 , ISSUE 4, 122–124

Article | 16-November-2020

A second example of anti-Esa, an antibody to a high-incidence Cromer antigen

A blood sample contained an antibody to a high-incidence antigen that reacted with all red blood cells (RBCs) tested by the indirect antiglobulin test (IAT). The antibody reacted with papain-, ficin-, and trypsin-treated RBCs, but not with α-chymotrypsin-treated RBCs. This pattern of reactivity suggested the possibility that the antibody was recognizing an antigen in the Cromer blood group system. Tests against RBCs deficient in decay-accelerating factor (which carries the Cromer antigens

Marion E. Reid, Roselyn Marfoe, Anita Mueller, Patricia A. Arndt, Laima Sausais, Peggy Spruell

Immunohematology, Volume 12 , ISSUE 3, 112–114

Article | 14-December-2020

The sensitivity of antibody detection testing using pooled versus unpooled reagent red cells

. Little data, however, have been published to support this contention. In the present study, the data show a decreased sensitivity for antibody detection when pooled reagent RBCs are used. This reduced sensitivity could result in failure to detect some clinically significant RBC alloantibodies, which might result in the occurrence of overt hemolytic transfusion reactions, especially if an indirect antiglobulin test is not performed at the time blood is crossmatched.

Ira A. Shulman, Roland Nakayama, Cintia Calderon

Immunohematology, Volume 7 , ISSUE 1, 16–19

Article | 14-October-2020

Evaluation of a new solid-phase immunoassay for alloantibody detection using bromelin-treated and untreated red blood cells

detected with enzyme-treated intact RBCs and untreated RBCs by M-MPHA. The slight increase in reactivity using M-MPHA was not seen using dried RBC stroma (M-MPHA-Dry). All donor-derived IgG alloantibodies, which were detected by either a conventional tube enzyme test or an indirect antiglobulin test, were detected by M-MPHA without using enzyme-treated RBCs. Both M-MPHA and M-MPHA-Dry can be used for antibody detection without using enzyme-treated RBCs and are also useful for antibody identification.

Toyohiro Tamai, Toshio Mazda

Immunohematology, Volume 17 , ISSUE 1, 17–21

Article | 14-December-2020

Antibody detection using pooled sera and a solid phase system

The purpose of the study was to evaluate the feasibility of substituting the Immucor Capture™-R solid phase (SP) antibody detection system for our routine donor antibody screen. Our routine procedure (RP) used a 12-drop pool of six donor sera and one drop of pooled reagent red cells, with 37°C incubation and indirect antiglobulin test readings. The SP system was used according to the manufacturer’s directions except that one drop of the pooled sera (rather than an individual

Malcolm L. Beck, Jill T. Hardman, Alicia M. Briseño

Immunohematology, Volume 7 , ISSUE 3, 73–75

Article | 18-October-2020

Human anti-Dia monoclonal antibodies for mass screening

The use of monoclonal antibodies (mabs) to blood group antigens is constantly increasing for routine typing. Two heterohybridoma cell lines, HMR15 and HMR22, were established by Epstein-Barr virus transformation of peripheral blood lymphocytes from a blood donor with anti-Dia. HMR15 mab directly agglutinated Di(a+) red cells, and HMR22 mab agglutinated Di(a+) red cells exclusively by the indirect antiglobulin test. Reactivities of both HMR15 and HMR22 mabs were specific for Dia and had good

Toru Miyazaki, Shinichiro Sato, Toshiaki Kato, Hisami Ikeda

Immunohematology, Volume 16 , ISSUE 2, 78–81

Article | 30-November-2020

An example of anti-LWa in a 10-month-old infant

Blood samples from a 10-month-old male infant requiring transfusion were found to contain an allomtibody reacting at 37°C in saline, by indirect antiglobulin test (IAT), and with a manual polybrene technique. Preliminary results suggested anti-D and another weaker reacting antibody, but the patient had been previously transfused with only D- blood. His serum reacted more weakly by IAT against red cells treated with 0.2M dithiothreitol (DTT), and one D+, LW(a-) sample was nonreactive. The

Alan Devenish

Immunohematology, Volume 10 , ISSUE 4, 127–129

Article | 10-November-2020

The second example of Lu:-7 phenotype: serology and immunochemical studies

We describe the second example of red blood cells (RBCs) with the Lu:–7 phenotype in a 37-year-old Latino female (SA). Her RBCs were nonreactive with anti-Lu7 (Mrs. GA) but were reactive with all other antibodies to high-prevalence antigens tested, including those in the Lutheran blood group system. No Lu:–7 RBCs were available for testing. SA’s serum was nonreactive by the indirect antiglobulin test against (1) recessive and dominant Lu(a–b–) RBCs and (2) trypsin

Marion E. Reid, Jack L. Hoffer, Ragnhild Øyen, Edith Alicea-Tossas, Manijeh Sadjadi, Gladys M. Messina

Immunohematology, Volume 12 , ISSUE 2, 66–68

Case report | 01-December-2019

A case of masquerading alloantibodies:  the value of a multitechnique approach

, D+ blood type showing strong reactivity with all cells tested  in the forward and reverse ABO, in the D testing as well as in a three-cell antibody screen. The initial assumption was that the plasma contained a cold autoantibody. Subsequent testing, including the use of gel column technology, ficin-treated cells, and antisera for phenotyping, showed the apparent cold autoantibody to be a red herring. Additional tube testing at immediate spin, 37°C, and indirect antiglobulin test (IAT

Paula M.S. Wennersten, Laurie J. Sutor

Immunohematology, Volume 30 , ISSUE 3, 117–120

Article | 22-November-2020

Use of monoclonal Jka and Jkb reagents in phenotyping red cells with a positive direct antiglobulin test

an indirect antiglobulin test. The Jka and Jkb MAbs consistently gave the same phenotype results both on untreated DAT-positive red cells and on the same cells after CDP treatment. Two of the CDPtreated samples had diminished antigen expression with the MAbs, a finding that may have been caused by the CDP treatment. One untreated sample, which spontaneously agglutinated in a lowprotein medium, was incorrectly phenotyped with the anti-Jka MAb, but both MAbs and PAbs gave the same correct results

Judy L. Brazell

Immunohematology, Volume 10 , ISSUE 1, 16–18

Case report | 09-October-2019

Hemolytic transfusion reaction attributable to anti-Dia  

In situations when a patient's antibody detection test is negative, many institutions have moved from an indirect antiglobulin test (IAT) crossmatch to an electronic crossmatch system. Here we report a case of an acute hemolytic transfusion reaction attributable to anti-Dia in a patient with a negative antibody detection test. A 22-year-old female patient with a diagnosis of β thalassemia and sickle cell anemia commenced a routine exchange transfusion of 5 units of red blood cells

Arthur J. Joyce, Kelli M Quantock, Ray Banh, Yew-Wah Liew

Immunohematology, Volume 33 , ISSUE 1, 6–8

Case report | 26-October-2019

Weak D type 67 in four related Canadian blood donors

Correct donor D typing is critical to prevent recipient alloimmunization. No method can detect all variants, and the immunogenicity of many variants is unknown. Routine ABO and D serologic typings are performed in our laboratory by automated microplate testing. Until 2011, routine confirmation of D– status of first-time donors was performed by the manual tube indirect antiglobulin test (IAT); this was replaced by automated solidphase testing including weak D testing by IAT. Selected

Philip Berardi, Emma Bessette, Michiko Ng, Nancy Angus, Debra Lane, Lise Gariepy, Katerina Pavenski, Gorka Ochoa-Garay, Jacqueline Cote, Mindy Goldman

Immunohematology, Volume 31 , ISSUE 4, 159–162

Article | 14-October-2020

Antibody screening in 37°C saline. Is it safe to omit it using the indirect antiglobulin (gel) test?

Pretransfusion tests must detect antibodies that can shorten the life of red blood cells (RBCs). Some studies have demonstrated the existence of clinically significant antibodies detected at 37°C in saline that are not detected by the indirect antiglobulin test (IAT) when the conventional tube test is used. Our aim was to determine whether these antibodies, detected with a 37°C saline tube test, are also detected when a sensitive column gel agglutination method is used. The 2373

José A. Duran, Manuel Figueiredo

Immunohematology, Volume 18 , ISSUE 1, 13–15

Article | 18-October-2020

Comparison of tube and gel techniques for antibody identification

There are several methods for antibody detection and each technique has advantages and limitations. We compared the performance of the tube (polyethylene glycol–indirect antiglobulin test [PEG-IAT]) and gel test technique for antibody identification. From January to May 1999, we performed antibody screening tests by gel and tube techniques on 10,123 random blood samples submitted to our reference laboratory. Six hundred and twentyeight (6.2%) reactive samples were tested for antibody

Marcia Cristina Zago Novaretti, Eduardo Jens Silveira, Edio da Costa Filho, Pedro Enrique Dorlhiac- Llacer, Dalton de Alencar Fischer Chamone

Immunohematology, Volume 16 , ISSUE 4, 138–141

Article | 26-October-2020

Anti-Lu9: the finding of the second example after 25 years

The first and only reported exanipie of anti-Lu9 (an antibody directed at a low-incidence antigen in the Lutheran blood group system and allelic to the high-incidence antigen Lu6) was described in 1973 in the serum of a white female, Mrs. Mull. Her serum also contained anti-Lul1 (-Lua), and subsequently, an anti-HLA-B7 (-Bga) was identified. We report the second example of anti-Lu9 in a white male (GR), found 25 years later. The GR serum was reactive in the indirect antiglobulin test with Lu

Kayla D. Champagne, Marilyn Moulds, Jo Schmidt

Immunohematology, Volume 15 , ISSUE 3, 113–116

Article | 17-November-2020

Analysis ofthe routine use of polyethylene glycol (PEG) as an enhancement medium

This study compared the performance of polyethylene glycol (PEG) and low-ionic saline solutions (LISS) as enhancement media for routine use in a large transfusion service. A PEG additive solution (PEG plus LISS) was compared to a LISS additive (LISS plus polymers) and to an albumin-indirect antiglobulin test (A-IAT). Fifty serum samples containing clinically significant alloantibodies and fifty samples without alloantibodies were tested. Following an acute hemolytic transfusion reaction (HTR

Vicki J. Barrett, James R. Stubbs, Karen Stuardi, Angela Hollis, Leslie Clear

Immunohematology, Volume 11 , ISSUE 1, 11–13

Article | 14-October-2020

Warm autoimmune hemolytic anemia with mimicking anti-c and -E specificities

An 18-month-old male was admitted to a hospital with a hemoglobin of 4.1 g/dL and a reticulocyte count of 53 percent. There was no history of prior transfusion. Serologic evaluation revealed the presence of both a positive direct antiglobulin test (DAT) and an indirect antiglobulin test (IAT). The patient’s red blood cells (RBCs) typed as group A, C–D–E–c+e+ (cde/cde). Evaluation of the IAT revealed the presence of anti-c and anti-E. All other major antibodies were ruled

Hsin-Yeh Hsieh, Diana L. Moroney, Deanne E. Naumann, D. Jane Hata, Nancy C. Vosnidou, Rovenna L. Kessinger, Nassir Shahab, Nasrollah Hakami, Daniel S. Smith

Immunohematology, Volume 18 , ISSUE 1, 19–22

Article | 17-February-2021

Prevalence of DEL phenotype in D– blood donors in India

samples negative for D by the indirect antiglobulin test (IAT) were found to be positive by adsorption-elution.3 Until recently, DEL was primarily described in Asians, since a significant proportion of D– Asian individuals are in fact DEL+.5–7 In Caucasians, DEL is very rare, although recent molecular studies show that the prevalence of DEL in donors typed D– in routine testing is greater than initially thought.8,9 Molecular studies have shown that a heterogeneous array of variant RHD alleles can

R. Chaudhary, S. Verma, A. Verma

Immunohematology, Volume 36 , ISSUE 4, 133–136

Article | 10-November-2020

Do monocyte ADCC assays accurately predict the severity of hemolytic disease of the newborn caused by antibodies to high-frequency antigens?

Monocyte ADCC assays are helpful indicators of the severity of hemolytic disease of the newborn (HDN) due to anti-D. It would be particularly useful if the assays also accurately predicted the ability of antibodies to high-frequency antigens (HFA) to cause HDN. To investigate this possibility, 14 antenatal sera containing antibodies to HFA were tested and the results correlated with the severity of HDN. Antibody titers were determined using an indirect antiglobulin test (IAT). Eight sera 

Stephen F. Garner, Alan Devenish

Immunohematology, Volume 12 , ISSUE 1, 20–26

Case report | 01-December-2019

Paroxysmal cold hemoglobinuria: a case report

Scott C. Wise, Sheila H. Tinsley, Lloyd O. Cook

Immunohematology, Volume 28 , ISSUE 4, 118–123

Report | 01-December-2019

Identifying D-positive donors using a second automated testing platform

estimated the number of cases of alloimmunization in women younger than 50 years likely to be prevented by the addition of Galileo testing. From May 2011 to May 2012, 910,220 donor samples were tested; 15,441 were first-time donors with concordant D– results. Five donors tested D– on the PK7300 and weak D+ on the Galileo; one was found to be a false positive on further testing. On manual testing, the other four donors had positive indirect antiglobulin test results with one to three of the

Mindy Goldman, Ilona Resz, Jacqueline Cote, Gorka Ochoa, Nancy Angus

Immunohematology, Volume 29 , ISSUE 3, 97–100

Article | 03-November-2020

Use of LOR-15C9 monoclonal anti-D to differentiate erythrocytes with the partial DVI antigen from those with other partial D antigens or weak D antigens

Historically, red blood cells (RBCs) with partial D antigens have been defined serologically by their pattern of reactivity with polyclonal and monoclonal anti-D. Although numerous variants have been described in tests with well-characterized monoclonal anti-D, definition remains difficult to ascertain serologically. RBCs of known partial D type were tested with LOR-15C9 (a monoclonal anti-D) and commercial anti-D by the tube indirect antiglobulin test (IAT), by micro typing system IgG gel

Marion E. Reid, Gregory R. Halverson, Francis Roubinet, P.A. Apoil, Antoine Blancher

Immunohematology, Volume 14 , ISSUE 3, 89–93

Article | 22-November-2020

Cefotetan-induced immune hemolytic anemia due to the drug-adsorption mechanism

epinephrine, and she was transfused with an additional unit of RBCs. Her Hct rose to 34.1% prior to her discharge on day 11. Further investigation revealed that the patient's serum and an eluate contained an antibody that reacted with cefotetan- and cephalothin-coated RBCs by the indirect antiglobulin test. In a monocyte monolayer assay, monocytes readily phagocytized cefotetan- and cephalothin-coated reagent RBCs but not uncoated reagent RBCs. The patient's serum did not react by the so-called

Robert J. Eckrich, Susan Fox, Delores Mallory

Immunohematology, Volume 10 , ISSUE 2, 51–54

Article | 30-November-2020

Misidentification of anti-Vel due to inappropriate use of prewarming and adsorption techniques

Two units of red blood cells (RBCs) were ordered for a 44-year-old Caucasian woman with renal failure and cancer. Pretransfnsion testing performed at the regional reference laboratory had revealed the presence of an antibody reactive with all cells at the indirect antiglobulin test (IAT) but apparently nonreactive by a prewarmed IAT. The patient’s RBCs were direct antiglobulin test negative. Adsorption of the serum with rabbit erythrocyte stroma or with allogeneic RBCs at 4°C reduced

Jill Storry, Delores Mallory

Immunohematology, Volume 10 , ISSUE 3, 83–86

Article | 06-December-2020

Failure to detect a prozoning anti-Fya in the serum of a chronically transfused patient

A 78-year old white male presented to our facility with vascular occlusion. He was Group A, D-positive, with a negative indirect antiglobulin test (antibody screen). Records at our facility, 4 years prior to this admission, indicated a history of anti-Fya, and Fy(a-) units were provided. A referring hospital had transfused the patient for "chronic anemia" (1-3 units weekly for 2 years), and he had received eight units (untested for Fya) immediately prior to his transfer to our

Jody C. Sizemore, Teresa D. Sammons, Jerry N. Clanton

Immunohematology, Volume 8 , ISSUE 2, 44–46

Article | 03-November-2020

Improved detection of weak, clinically significant antibodies by supplementation of polyethylene glycol with a low-ionic solution

A comparative study of 164 serum samples was carried out to determine the specificity and sensitivity of the indirect antiglobulin test (IAGT) in three different formulations: physiologic saline, low-ionic solution (RAM), and RAM supplemented with polyethylene glycol (PEG). Serum samples containing mostly weak antibodies (anti-D, -C, -E, -c, -Jka, -Fya, -K, -S, -Lea, -Lua, -M, -Cob, -P1, -I, and -Kna) were used in a 10-minute IAGT in which PEG-IAGTs were compared with salineIAGTs and RAM-IAGTs

Kim Swee Low, Yew-Wah Liew, Peter M. Bradley

Immunohematology, Volume 14 , ISSUE 2, 68–71

Article | 22-January-2021

Routine indirect antiglobulin testing of blood donors—a further step toward blood safety: an experience from a tertiary care center in northern India

transfusions, since the presence of unexpected antibodies in the donor plasma can lead to adverse transfusion reactions, particularly in cases in which a large amount of plasma is transfused (as in massive transfusions) and in pediatric patients.3 In this scientific article, we emphasize the importance of establishing a policy for antibody screening of all blood donors as a step to further improve blood safety. The policy of routine inclusion of the indirect antiglobulin test (IAT) was adopted in our

S. Malhotra, G. Negi, D. Kaur, S.K. Meinia, A.K. Tiwari, S. Mitra

Immunohematology, Volume 36 , ISSUE 3, 93–98

Report | 01-December-2019

Comparison of estimation of volume of fetomaternal hemorrhage using KleihauerBetke test and microcolumn gel method in D-negative nonisoimmunized mothers

In this study we assessed the efficacy of the microcolumn gel method in the detection and quantification of the volume of fetomaternal hemorrhage (FMH) in comparison with the Kleihauer-Betke test (KB) in nonisoimmunized D– mothers. We collected blood samples from 80 D– indirect antiglobulin test–negative mothers over a span of more than 1 year. FMH was determined by KB and microcolumn gel method, and the results were compared. FMH was recorded as less than 4 mL by KB if no

Kshitija Mittal, Neelam Marwaha, Praveen Kumar, Subhash C. Saha, Beenu Thakral

Immunohematology, Volume 29 , ISSUE 3, 105–109

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