Identification of antibody against highly prevalent antigen through serological test and molecular biology technology / 中国输血杂志

【Objective】 To identify a case of antibody against highly prevalent antigen through molecular biology technology. 【Methods】 Blood group typing, unexpected antibody identification and cross matching were performed by serological test, and genetic testing of Diego blood group was performed by molecular biology technology. 【Results】 Serological test showed that there was a high prevalence of anti-Dib in the serum of the patient. Gene sequencing showed that the genotype of the patient was Di(a+b-) . Two cases with Di(a+b-) matched with the patient were screened from 856 blood donors. 【Conclusion】 The combined detection method based on serological test supplemented by molecular biology technology is beneficial to the detection of antibody against highly prevalent antigens, and is of great significance for ensuring the safety of clinical blood transfusion.

Unexpected antibody distribution among tumor patients: analysis of 111 483 cases / 中国输血杂志

【Objective】 To analyze the distribution of unexpected antibodies in tumor patients retrospectively and explore the clinical significance. 【Methods】 Unexpected antibody screening was performed on inpatients with blood preparation and blood transfusion in our hospital from January 2004 to December 2022, with 1 176 cases tested positive, and the types of unexpected antibodies and distribution characteristics were statistically analyzed. 【Results】 Unexpected antibodies were screened in 1 176 cases, with the positive rate at 1.05% (1 176/111 483). The unexpected antibodies were mainly anti-E 16.33%(192/1 176), anti-M 7.99% (94/1 176), anti-Mur 5.70% (67/1 176) and anti-Lea 4.76% (56/1 176). Among the 1 176 cases, gastrointestinal tumors accounted for 27.99% (329/1 176), gynecological tumors accounted for 24.84% (292/1 176), respiratory tumors accounted for 16.67% (196/1 176) . 【Conclusion】 The influencing factors of unexpected antibodies in tumor patients were disease type, blood transfusion history and blood type. Therefore, it is necessary for clinical departments to carry out unexpected antibody screening and perform Rh blood type matched transfusion for tumor patients to avoid alloantibody production.

Identification of Complex and Combined Antibody Consisted of Anti-c, Anti-E, Anti-Jka and Anti-Fya / 中国实验血液学杂志

OBJECTIVE@#To investigate the role of multiple serological methods in the identification of complex antibodies.@*METHODS@#The blood group antigens were detected by saline and microcolumn agglutination methods. The saline method was used to screen and identify IgM-type antibodies in the patient's serum, while the polybrene, anti-globulin, microcolumn agglutination, enzymic and absorption-elution methods were used to screen and identify IgG-type antibodies.@*RESULTS@#The patient was B/CCDee/Jk(a-b+)/Fy(a-b+) blood type. The serum reacted with panel cells, and the reaction presented anti-E pattern in the saline medium. It was fully positive in the microcolumn agglutination card, except 2 negative ones after using papain to treat the panel cells. Referring to the pattern table, it was concluded that there existed anti-c, anti-E, and anti-Jka antibodies, and one antibody corresponding to an antigen that was easily destroyed by papain. The red blood cells with specific phenotype were selected for absorption-elution to identify IgG-type anti-c, anti-E, anti-Jka and anti-Fya antibodies.@*CONCLUSION@#It is confirmed that IgM-type anti-E, and IgG-type anti-c, anti-E, anti-Jka and anti-Fya antibodies exist in the patient's serum by multiple serological methods.

How to eliminate the interference of warm autoantibodies with antibody identification / 中国输血杂志

【Objective】 To investigate the effect of serological methods for eliminating the interference of warm autoantibodies with the compatibility test before blood transfusion. 【Methods】 The blood samples (whole blood and serum, 3 mL/each) of 10 cases of warm autoantibodies interfering with antibody screening and identification were collected. Autogenous or allogeneic red blood cells(RBCs) were treated with microthermal(45 ℃), chloroquine, or ZZAP (dithiothreitol and papain) reagents, and then were used to absorb the autoantibodies in patients′ plasma. The plasma after absorption and RBC elution were used for antibody identification by anti-human globulin method or Polybrene method to determine the specificity of the autoantibody/alloantibody. Blood transfusion with ABO/Rh homotypic RBCs without corresponding antigens was performed in patients with alloantibodies or specific autoantibodies, and the efficacy of blood transfusion was evaluated. 【Results】 The interferenc of warm autoantibodies with antibody screening and identification due to primary or secondary autoimmune diseases were eliminated after absorption, and IgG isospecific antibodies (anti-E, anti-Jka, anti-Wra) in 3 cases and specific autoantibodies (anti-Ce) in 1 case were yielded. 6 of the 10 patients received blood transfusion, and 4 with specific antibodies avoided exposure to corresponding antigens. After transfusion of 2U suspended RBCs, the increase of hemoglobin at 24h in all 6 patients was greater than 10 g/L, and no hemolytic transfusion reaction occurred. Anemia symptoms were improved after transfusion. 【Conclusion】 Appropriate elution methods and autologous/allogeneic absorption methods can eliminate the interference of warm autoantibodies with the identification of alloantibodies, therefore can accurately identify the types and properties of antibodies, thus providing targeted and effective blood transfusion.

Approaches against the interference of autoantibodies with pre-transfusion compatibility testing / 中国输血杂志

Pre-transfusion compatibility testing is complicated in autoimmune hemolytic anemia (AIHA) patients due to the presence of autoantibodies. Delays in blood transfusion or even life-threatening would occur if blood type, isoantibodies/ autoantibodies of these patients could not be correctly identified to choose the appropriate blood components. Knowing the detection and treatment countermeasures against blood transfusion compatibility in AIHA patients is of great significance to ensure the timeliness and safety of blood transfusion. Based on the research progress at home and abroad, this article summarizes the serological characteristics, autoantibody types, blood group identification methods, antibody screening and antibody identification methods, and blood transfusion strategies about AIHA patients, in order to eliminate the interference of autoantibodies and provide transfusion guidance for the staff of Blood Transfusion Department.

Expert consensus of identification of antibodies to red cell antigens / 中国输血杂志

Identification of antibodies to red cell antigens is one of the important contents of blood transfusion compatibility test. The purpose is to accurately identify the alloantibodies in patients, and assess its association with fetal and neonatal hemolytic disease, hemolytic transfusion reaction, or a notable decrease in transfused red blood cell survival. This consensus summarizes the key points of quality management, requirements for samples and assay, principle and selection of detection methods, process of routine antibody identification, interpretation of result and comprehensive analysis and interpretation of erythrocyte blood group antibody identification, aiming to improve the correct understanding of the significance of antibody identification in the lab and guarantee the sensitivity and accuracy of antibody identification test.

Hemolytic disease of newborn caused by multiple Rh antibodies

Hemolytic disease of Fetus and Newborn (HDFN) usually results due to natural occurring antibodies or alloimmunization in mother but the presence of multiple red cell antibodies increases the risk of development of significant HDFN. Here author reported a case of hemolytic disease of fetus and newborn in a preterm baby caused by multiple maternal antibodies. Direct Antiglobulin Test (DAT) on neonate blood sample was positive (3+) with monospecific DAT showed IgG type which was confirmed by heat elution. Antibody identification of eluate was done using commercial 11-cell panel by gel method showing specificity to anti-D and anti-C antibody which was differentiated from anti-G by sequential adsorption and elution studies. Neonate was treated with double volume exchange transfusion (DVET) using leucoreduced, irradiated O Rh D and C negative PRBC suspended in AB plasma and discharged 6th day in a stable condition. So, all pregnant women should be at least advised for ICT irrespective of Rh D negative status. If ICT is positive, they should be referred to higher center for proper Immunohematological work up, so that proper blood unit for DVET could be identified.

Clinical significance of Rh antigen typing detection for patients who require blood transfusions repeatedly / 中国免疫学杂志

Objective:The Rh antigen typing and antibody identification results of the patients whose irregular antibody screening positive were analyzed,to explore the clinical significance of detecting the Rh antigens typing before the first blood transfusion of the patients who require multiple blood transfusions.Methods:The Rh antigenic typing of 128 irregular antibody screening positive patients were tested by test tube method.The monospecific antibody were identified by microcolumn gel method.Results: Of the 128 patients with irregular antibody screening positive,there were 77 cases in Rh system,including 72 cases of anti-E and 5 cases of anti-c. There were 14 cases of MNSs system,including 10 cases of anti-M and 4 cases of anti-Mur.There were 15 cases of anti-Leain Lewis system.There were 4 cases of anti-P1in P system and 18 cases of other nonspecific antibodies.The distribution of Rh antigen detection was DCCee(74 cases)> DCcEe(34 cases)> DCcee(10 cases)> DccEE(5 cases)> DccEe(2 cases)> DCcEE(1 case),dCcee(1 case),dccee(1 case).The majority phenotype of Rh system antibodies in patients were DCCee.The patients were mainly distributed in the wards who require repeated blood transfusions such as the department of blood internal medicine(26 cases),digestive internal medicine(11 cases),ICU(4 cases).Conclusion:Before the first blood transfusion,we detect the Rh antigenic typing and choose the same antigen phenotype of Rh system for the patients who require blood transfusions repeatedly,which can avoid producing the irregular antibodies in this system,and then to ensure the safety and effective of the blood transfusion.

A Case Report of Anti-f(ce) Identified in a Patient with Pancreatic Cancer / 대한수혈학회지

Anti-f(ce) is a rare unexpected antibody against the ce(f) antigen. The aim of this study is to report a second case of anti-f(ce) identified in a patient. A 66-year-old-male with pancreatic cancer received percutaneous transhepatic biliary drainage. During pretransfusion tests, anti-f(ce) was identified. He had a history of multiple transfusions and was transfused with 2 units of antiglobulin crossmatch compatible RBCs without any adverse reactions. To confirm that the antibody was specific for ce(f) antigen, we crossmatched the patient's serum with RhD-positive red cells of Rh phenotype DcE, DCcEe, DCce, and DCe; all results were negative. Conversely, a crossmatch with RhD-negative red cells of Rh phenotype ce, Cce, and cEe, showed positive results for Rh phenotype ce and cEe red cells. Among the four reports that confirmed anti-e, we discovered the possibility of co-existence of anti-C or misidentification of anti-Ce as anti-e. Therefore, when antibodies against Rh antigens are identified, the possibility of co-existence of antibodies against compound antigens should be considered. Using unexpected antibody identification panel that ce(f) antigen positive red cells are marked is recommended for sensitive detection of anti-f(ce).

The Frequency of Unexpected Antibodies and Clinical Characteristics of Transfusion Candidates in a General Hospital During the Past 12 Years / 임상검사와정도관리

BACKGROUND: Unexpected antibodies are an important cause of hemolytic transfusion reaction. Thus, unexpected antibody screening and identification tests should be performed before every transfusion. We evaluated the frequency and distribution of unexpected antibodies and the clinical characteristics of patients in a Korean secondary general hospital with 745 beds in the past 12 years. METHODS: Between March 2000 and October 2011, unexpected antibody screening and an identification test using the Bio-Rad ID-System (Bio-Rad, USA) were performed in 72,600 patients. RESULTS: Of the 72,600 patients, 467 (0.64%) showed positive results for antibody screening tests. Among them, alloantibodies were identified in 324 (69.4%) patients and the types of alloantibodies were not identified in 64 (13.7%) patients. Autoantibodies were detected in 71 (15.2%) patients and panagglutination reactions were detected in 8 (1.7%). Of the 467 cases, 164 (35.1%) had a history of transfusion in our hospital. Among the 324 patients in whom alloantibodies were identified, anti-E (37.3%), anti-Lea (16.7%), anti-E and anti-c (14.8%), anti-C and anti-e (5.6%), anti-Leb (4.9%), anti-D (4.6%), anti-Jka (3.1%), anti-S (2.5%), and anti-M (1.9%) were detected. In 41 of the 324 (12.7%) of these patients, the types of antibodies were identified with the NaCl/Enzyme gel test but not with the LISS/Coombs gel test. CONCLUSIONS: Among the 467 patients, 130 (27.8%) in whom unexpected antibodies were detected, were scheduled for surgery. Because 101 of these 130 patients (77.7%) were unimmunized, unexpected antibody screening may be important in secondary hospitals with patients who do not have a detailed transfusion history. We identified Rh, P, and Lewis group antibodies more efficiently with a combination of the LISS/Coombs gel test and the NaCl/Enzyme gel test.

Comparison of Cryopreservation Methods of Rare Red Blood Cells Used for Antibody Identification Tests / 대한수혈학회지

BACKGROUND: When a certain antibody, which is not clearly identified, is detected before transfusion, rare red blood cells (RBCs) should be used for antibody identification. But its rarity make it difficult to identify the antibody. This study compared the high glycerol method with the liquid nitrogen method for the cryopreservation of rare RBCs that are used for antibody identification. METHODS: The frozen RBCs were thawed every 2 months to measure the strength of agglutination. We observed the changes of the strength of agglutination. The Di(a) antigen, which is relatively frequent in Asians, was included in this study. RESULTS: Using the high glycerol method, the decrease of the strength of agglutination was observed with using the k antigen from 4 months and the decrease of the strength of agglutination was observed with using M, N, s, Le(a), Le(b) and Fy(b) antigens from 8 months. With freezing the donor RBCs by the liquid nitrogen method, the decrease of the strength of agglutination was observed with using the C, s, k and Le(b) antigens from 2 months, with using the M and S antigens from 4 months, with using the D, c, e, N and Fy(a) antigens from 6 months and with using the Le(a) antigen from 8 months. Rare RBCs with the Di(a) antigen were successfully cryopreserved for 6 months with using the high glycerol method. For the commercial screening cells, the high glycerol method showed effective cryopreservation with using c, e, M, k, Le(a), Le(b), Jk(a) and Fy(b) antigens for 6 months or longer. CONCLUSION: We were able to preserve most of the important antigens of the RBCs for at least 6 months without a significant decrease of reactivity by employing the high glycerol cryopreservation technique.

Comparison of Cryopreservation Methods of Rare Red Blood Cells Used for Antibody Identification Tests / 대한수혈학회지

BACKGROUND: When a certain antibody, which is not clearly identified, is detected before transfusion, rare red blood cells (RBCs) should be used for antibody identification. But its rarity make it difficult to identify the antibody. This study compared the high glycerol method with the liquid nitrogen method for the cryopreservation of rare RBCs that are used for antibody identification. METHODS: The frozen RBCs were thawed every 2 months to measure the strength of agglutination. We observed the changes of the strength of agglutination. The Di(a) antigen, which is relatively frequent in Asians, was included in this study. RESULTS: Using the high glycerol method, the decrease of the strength of agglutination was observed with using the k antigen from 4 months and the decrease of the strength of agglutination was observed with using M, N, s, Le(a), Le(b) and Fy(b) antigens from 8 months. With freezing the donor RBCs by the liquid nitrogen method, the decrease of the strength of agglutination was observed with using the C, s, k and Le(b) antigens from 2 months, with using the M and S antigens from 4 months, with using the D, c, e, N and Fy(a) antigens from 6 months and with using the Le(a) antigen from 8 months. Rare RBCs with the Di(a) antigen were successfully cryopreserved for 6 months with using the high glycerol method. For the commercial screening cells, the high glycerol method showed effective cryopreservation with using c, e, M, k, Le(a), Le(b), Jk(a) and Fy(b) antigens for 6 months or longer. CONCLUSION: We were able to preserve most of the important antigens of the RBCs for at least 6 months without a significant decrease of reactivity by employing the high glycerol cryopreservation technique.

The Effect of Number of Reagent Red Cells on the Antibody Screening and Identification / 대한수혈학회지

BACKGROUND: The number of reagent red cells has an effect on the results of an unexpected antibody screening test. We evaluated the effect of the number of reagent red cells on antibody screening and identification using test panels of a DG Gel (Diagnostic Grifols, Barcelona, Spain). METHODS: A total of 310 samples were tested in parallel using SeraScan Diana 2 and SeraScan Diana 4 (Diagnostic Grifols, Barcelona, Spain) and ID-DiaCell (DiaMed, Cressier, Morat, Switzerland). Positive samples as determined by the use of SeraScan and ID-DiaCell were identified on an ID-Dia panel (DiaMed), Identisera Diana and Identisera Extend (Diagnostic Grifolsn). RESULTS: Among the 310 samples, 54, 59 and 59 samples were determined as antibody positive by the use of SeraScan Diana 2, SeraScan Diana 4 and ID-DiaCell, respectively. Unexpected antibodies were identified in 10/59 samples (17%) by the use of SeraScan Diana 4, and were identified in 34/59 samples (57.6%) by the combined use of SeraScan Diana 2 and SeraScan Diana 4. Identification of unexpected antibodies by the use of Identisera Diana or Identisera Extend was not different. CONCLUSION: When the results of SeraScan Diana 2 and SeraScan Diana 4 are integrated, unexpected antibodies could be identified in 57.6% of the screening-positive samples. Therefore, if the number of reagent red cells is increased, some antibodies can be identified by antibody screening tests, and the results can be used to validate those of antibody identification tests.

Diagnostic Usefulness of Antibody Identification Test Including Enzyme Method

BACKGROUND: Many medical institutions in Korea have recently been performing an antibody screening test as one of the essential elements of a pre-transfusion test. In this study we will determine the advantage and clinical significance of adding an enzyme method to the antiglobulin method while conducting the antibody identification test. METHODS: We performed antibody identification tests between December 2002 and December 2005, for a total of 37 months at Pusan National University Hospital. In this study we have analyzed 550 cases that were conducted by both the antiglobulin method and the enzyme method at the same time. RESULTS: A total of 111 of the results were cases of detection using the adding an enzyme method. Among these results, Rh antibodies that included the anti-E had the highest number of results 77 (69.4%), 28 antibody (25.2%), 2 anti-P1 (1.8%) and one anti-Jkb (0.9%). CONCLUSION: Using the enzyme method in the antibody identification test proved to us that there were more clinically significant warm antibodies than cold antibodies. In order to have a more secured transfusion, it is required to identify a clinically significant antibody using the additional enzyme method during the antibody identification test.

The Frequency of Unexpected Antibodies in Blood Donors and Transfusion Candidates in Korea / 대한수혈학회지

BACKGROUND: A retrospective study was performed to estimate the frequency of red cell antibodies in blood donors (n=1,620,023) and transfusion candidates (SNUH n=12,111, YUMH n=26,665) for last 2 years (2000~2001). The results of the antibody screening and identification tests, the frequency and specificities of antibodies identified compared with blood centers and two hospitals had been used the different test methods each others. METHOD: Blood centers had been used tube and micro-plate method simultaneously with an in house and commercial panels. SNUH had been used micro-plate method using V plate with an in house and commercial panels. YUMH had been used gel agglutination technique (DiaMed ID System : DiaMed, Murten; Switzerland) since 1998. RESULTS: The frequencies of red blood cell antibodies were 0.26% (4,204 / 1,620,023 donor sera ), 0.11% (135 / 12,111 patient sera in SNUH) and 0.48% (128 / 26,665 patient sera in YUMH). Female donors and old ages showed higher frequency of red cell antibodies than male and young ages. Most of antibodies detected in donors were clinically less relevant antibodies such as Anti-Lea and Leb (38.9%), anti-P1 (18.1%), anti-H(IH) (8.4%), anti-M (6.2%) and so on. Clinically significant antibodies including Rh system antibodies (2.0%) were few, and composed only 12% of all the antibodies detected. In patients, clinically relevant antibodies including Rh antibodies (40.4% in SNUH, 71.9% in YUMH) were more frequently observed comparing with in donors. CONCLUSION: Antibodies found in donors were mostly clinically less relevant. Antibody screening method used in blood centers would be standardized. Blood banks using gel technique showed high detection rate of clinically significant antibodies comparing with facilities using other methods.

Experience of HLA Antibody Testing in the International Serum Exchange Program / 대한진단검사의학회지

BACKGROUND: When organ transplantation or HLA-matched platelet transfusion is considered, accu-rate identification of HLA antibody specificity in the recipient's serum is very important. In this study, we report our experience in an international quality control program. METHODS: For external quality control in a HLA antibody test, the International Serum Exchange Program distributes serum samples, generally showing polyspecific reactivity for cross-reactive epitope groups (CREGs), to participating laboratories: 4 samples per survey, 10 surveys per year. Participating in the program from May 1998 to August 2000 (24 surveys), we performed HLA antibody identification of 96 serum samples by the AHG-CDC (anti-human globulin-complement dependent cytotoxicity) method using frozen lymphocyte trays (36 lymphocyte panels). We compared the results of our laboratory with those of the total participants (all methods combined, 72 to 92 laboratories per survey) using the analyzed survey results distributed by the program organizer. RESULTS: We analyzed the survey results for the antibodies to relatively common HLA antigens in Koreans (antigen frequency >1%). For the HLA antibodies detected in >or=20% of participants, our detection rate was higher by 10-15% than that of all laboratories (HLA-A, 76% vs 65%; HLA-B, 73% vs 57%). And for the HLA antibodies detected in >or=50% of the participants, our detection rate was as high as 88% for HLA-A and 87% for HLA-B. Our detection rate for a few antibody specificities was lower than that of all laboratories, namely HLA-A1, A3, B35, and B55. Among these, A1, A3, and B55 were of lower incidence antigens in Koreans (antigen frequency 3-4%), indicating that the low detection rate was due to a limitation in the composition of lymphocyte panels. CONCLUSIONS: In general, our detection rate of HLA antibodies was superior to the average detection rate of the total participant laboratories. We would be able to improve the low detection rate for a few antibody specificities to lower incidence antigens by refining the composition of lymphocyte panels.

Usefulness of column agglutination test for irregular antibody screening and identification / 대한임상병리학회지

BACKGROUND: Type and screen is recommended for efficient use of blood and reduction in workload in blood bank. Column agglutiation test is standardized and easy to perform, and provide clear and stable reactions that improve the interpretation of results. In this study, we compared column agglutination test(Ortho Diagnostic Systems Inc., USA) with conventional tube test and investigate its usefulness in irregular antibody screening and identification. METHODS: A total 182 samples were screened for irregular antibodies using column agglutination test and conventional tube test. And 18 patient's sera in which irregular antibodies previously screened by both tube and column agglutination tests were identified for irregular antibodies by tube and column agglutination tests. We compared the results of two tests. RESULTS: In the screening test, there was 96.7%(176/182) agreements between column agglutination test and conventional tube test. The column agglutination test showed stronger reactivity than tube test. In the irregular antibody identification, there was 88.8%(16/18) agreement between two tests and disagreement were seen in the identification of anti-P1 and anti-Leb antibodies. CONCLUSION: The results of column agglutination test are objective and superior to the conventional tube test in irregular antibody screening and identification tests. These results suggest that the column agglutination test will be useful and more convenient test in antibody screening and identification.

Development of decision support system for antibody identification / 대한수혈학회지

BACKGROUND: Determination of antibody specificity using antigram spread sheet requires experience and knowledge on in vitro characteristics of red cell antibodies, time-consuming, and still subjective to human error. A computer-based antibody identification system was developed to overcome these disadvantages. METHODS: Decision support system program for antibody identification was designed using Visual Basic 5.0 for Dade Data-cyte Plus. This system integrates the reaction patterns of saline, 37degrees C albumin, antiglobulin, 4degrees C saline enzyme treated and user-defined phases and lists the antibodies according to the probability. 115 irregular antibodies previously confirmed by standard manual method reanalyzed with this program. RESULTS: In 111 of 115 cases (96.5%), this system produced the same results with the manual identification. In two cases, of not matched 4 cases the computer program suggested additional antibodies and in one case, the computer program detected previous human error. In the other case, antibody identification was possible only after further tests including selective adsorption of multiple antibodies. CONCLUSION: The decision support system was rapid and easy and showed good concordance rate when compared with manual antibody identificaion results. In addition, human error could be reduced. Decision support system for antibody identification could be used in small blood banks by less experienced staffs.