BACKGROUND: Since the 1960 s rabbit antihuman globulin reagent has been used widely. Recently most conjugate of enzyme immunoassay is produced from goat, and precise purification method is developed. Therefore we evaluated the commercial value of the goat antihuman globulin as a blood bank test reagent. METHODS: The human IgG was purified by protein-A gel, and injected into goat. The goat antihuman globulin was coupled by CNBr activated sepharose 4B-gel and purified by 0.2M glycine elution buffer. For verification of this reagent, commercial reagents(Ortho rabbit reagent & DiaMed Gel test) were used. RESULTS: The minimal concentration for detecting antibody of goat reagent was 9 ng/mL. The results of direct antiglobulin tests, with 400 samples collected from donated blood in CPDA-1, were all negative(false positive rate: 0%). With 613 samples collected from inpatients of Severance Hospital, the results were positive in 35 patients(positive rate:5.7%), and those results were in complete agreement with commercial reagent(concordance rate: Goat vs. Ortho :99.8%, Goat vs. DiaMed :98.4%). And with 30 samples of artificially prepared complement-coated RBC, the results were all negative. Indirect antiglobulin test showed higher agglutination score than commercial reagents. CONCLUSIONS: Goat reagent showed high sensitivity and specificity in comparison with rabbit reagent. Because false positive reaction was not observed in negative control samples, the heterophil agglutinin reaction, which was the major problem when the reagent was initially developed, might be excluded. In conclusion, goat reagent seems to be more economical than rabbit reagent because the former can be obtained in a large quantity and of high potency.
Agglutination ; Blood Banks ; Coombs Test ; False Positive Reactions ; Glycine ; Goats* ; Humans ; Immunoenzyme Techniques ; Immunoglobulin G ; Indicators and Reagents ; Inpatients ; Sensitivity and Specificity ; Sepharose

BACKGROUND: Major ABO incompatibility between donor and recipient is not a barrier for successful bone marrow transplantation(BMT). However, isoagglutinins could cause serious adverse effects, such as engraftment failure, delayed hematopoiesis and delayed hamolysis. It is important to detect these complications and bone marrow engraftment at an early stage for appropriate management. We have investigated the usefulness of isoagglutinin titer in major ABO incompatible BMT. METHODS: During April, 1996 to September, 1997, thirteen cases underwent major ABO-incompatible BMT at Asan Medical Center. We reviewed their medical records and performed ABO blood typing, IgM and IgG isoagglutinin titration, direct antiglubulin test and test for A and B transferase activities. RESULTS: Isoagglutinins against the donor antigen disappeared in 9 cases, decreased in 3 cases and persisted in 1 case. One patient who had persistent isoagglutinin titer had not had a bone marrow engraftment. The average time for anti-A isoagglutinin titer to disappear was 167.8 days(range; 90-281) and for anti-B was 116.8 days(range; 30-276). Eleven out of 13 cases, the RBCs of donor type appeared. On the average, RBCs of group A appeared later than RBCs of group B (112.7 +/- 99.6 days vs. 71.4 +/- 92.0 days, P>0.05). In 7 out of 8 cases that were done the A and B transferases test, the donordegrees empty set s RBC was produced but very little or none of the donor type transferase appeared in the serum. CONCLUSION: In order to determine the erythroid engraftment after the major ABO incompatible BMT, it is necessary to check the isoagglutinin titer periodically.
Blood Grouping and Crossmatching ; Bone Marrow Transplantation* ; Bone Marrow* ; Chungcheongnam-do ; Hematopoiesis ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Medical Records ; Set (Psychology) ; Tissue Donors ; Transferases

BACKGROUND: Currently brain dead solid organ transplantations are performed in several institutions, and these are extended in Korea. Especially liver transplantation requires such a large amounts of blood components including filtered and irradiated cellular components that blood bank should give a great support to provide them. For effective management and reducing workload of blood bank in solid organ transplantation, we evaluated the blood component usage according to the type of organ transplantation and suggest a guideline for its optimal blood ordering schedule. METHODS: From February 1995 to October 1997, 143 solid organ transplantations (18 adults and 7 pediatric liver transplants, 115 renal transplants and 3 heart transplants) were performed in Samsung Medical Center. We investigated amount of blood components requested by surgeons or anesthesiologists, and evaluated their usage, discard rate and C/T ratio (crossmatch to transfusion ratio) during perioperative, intraoperative and postoperative period for solid organ transplantation. In liver and heart transplantation, the usage of blood component according to the operative phases was also evaluated. RESULTS: All of the patients who underwent liver and heart transplantation and 15% of the patients who underwent renal transplantation were transfused with blood components during operation. For adult liver transplantation, 31.1 units of leukocyte-depleted red blood cells (LDRBC), 43.6 units of fresh frozen plasma (FFP) and 16.3 units of leukocyte depleted platelets (LDPC) on an average were transfused. Intraoperative salvage using Cell Saver was performed in liver transplantation and the volume of salvaged was 7127.6 mL which was equivalent to 28.5 units of RBCs. The C/T ratio of RBCs was 1.4. In pediatric liver transplantation, 4.8 units of LDRBC and 4.3 units of FFP were transfused with C/T ratio of 1.9. Two of 5 pediatric liver transplantation donors were transfused with 3 units of RBCs, 1.5 units of FFP and 1.0 unit of whole blood by preoperative autologous blood donation. Only 18 out of 115 patients who underwent renal transplantation were transfused with 2 units of RBCs and 2 units of FFP. The discard rate revealed over 60% and C/T ratio was 4.6-5.1 in renal transplantation. For the heart transplantation 1.3 units of RBCs, 5.6 units of FFP, and 7.3 units of LDPC were transfused. The C/T ratio was 3.8. CONCLUSION: Compared with foreign reports, slightly larger amount of blood components were used for liver transplantation, however similar amount were used for renal and heart transplantation. As the results of present study, we propose a guideline for optimal blood ordering schedule for solid organ transplantation considering the marginal safety : 40 units of LDRBC, 50 units of FFP, 20 units of LDPC and 8 units of Cryoprecipitate for adult liver transplantation; 5 units of LDRBC and 6 units of FFP for pediatric liver transplantation; 2 units of LDRBC, 6 units of FFP and 10 units of LDPC for heart transplantation. Additional requests of blood components for liver and heart transplantation might be decided considering the clinical situations.
Adult ; Appointments and Schedules ; Blood Banks ; Blood Donors ; Brain Death ; Erythrocytes ; Heart ; Heart Transplantation ; Humans ; Kidney Transplantation ; Korea ; Leukocytes ; Liver ; Liver Transplantation ; Organ Transplantation* ; Plasma ; Postoperative Period ; Tissue Donors ; Transplants*

BACKGROUND: Recently some countries such as U.S.A. and Canada where human immunodeficiency virus(HIV) infection is rather prevalent, included HIV-1 p24 antigen test as a routine donor blood screening. This study was performed to evaluate the advantage of additional p24 antigen testing for the prevention of transfusion-associated AIDS infection in Korea. METHODS: Blood collected from 1726 volunteer blood donors, 16 HIV-positive patients, 39 sera from 4 commercial seroconversion panels, 15 sera included in low titer performance panel were tested with HIV-1 p24 Antigen ELISA Test System(Ortho Diagnostic Systems, U.S.A.). Anti-HIV antibody was also measured in parallel employing commercial kits produced by two world-famous companies. For some sera, western blot testing was additionally done. RESULTS: False-positive rate of p24 antigen testing was 0.06%. In 2 examples from 4 seroconversion panels, the p24 antigen test detected HIV infection 1-25 days and 11-47 days earlier than anti-HIV tests. CONCLUSION: Additional p24 antigen testing was found to have a potential to reduce transfusion-associated HIV infections. Including the p24 antigen testing as a routine donor screening should be considered if the number of transfusion-associated HIV infections continues to grow in Korea.
Blood Donors ; Blotting, Western ; Canada ; Donor Selection ; Enzyme-Linked Immunosorbent Assay ; HIV Infections ; HIV-1 ; Humans* ; Korea ; Mass Screening ; Tissue Donors ; Volunteers

BACKROUND: Polymorphism of glycoprotein IIIa on human platelets is one of the factors in alloimmunization that causes neonatal alloimmune thrombocytopenia (NATP), and the granulocyte antigens NA1 and NA2 are often targets of granulocytes antibodies causing neonatal alloimmune neutropenia (NANP). Currently, serotyping relies on the properties of the typing sera or antibodies and technique used. Genotyping circumvents the problems associated with serotyping. METHODS: The genomic DNA of 200 unrelated pregnant women admitted to Taegu Fatima Hospital was typed for three platelet glycoprotein IIIa-specific antigens (HPA-1, HPA-4, and HPA-6w) and granulocyte antigens (NA1 and NA2). Allele specific amplification test using primer designed to study HPA-1 and HPA-4, restriction fragment length polymorphism to study HPA-6w, and sequence specific primers for NA1 and NA2 were used for genotyping. RESULTS: The genotype frequencies were HPA-1(a+b-) 100%, HPA-4 (a+b-) 97.5%, HPA-4(a+b+) 2.5%, HPA-6w(a+b-) 97%, and HPA-6w(a+b+) 3%. These frequencies are similar to Japanese but different from Caucasian. The gene frequencies of NA1 and NA2 were 0.56 and 0.44 respectively. There are no cases of alloimmune thrombocytopenia and neutropenia in newborns from the 200 studied women. CONCLUSIONS: The differences in genotype frequencies among platelet glycoprotein IIIa-specific antigens and in the gene frequencies of NA in Koreans are shown as compared with other ethnic groups. Therefore it is needed to find the proper screening target antigens and antibodies for Korean NATP and NANP patients.
Alleles ; Antibodies ; Antigens, Human Platelet ; Asian Continental Ancestry Group ; Blood Platelets* ; Daegu ; DNA ; Ethnic Groups ; Female ; Gene Frequency ; Genotype* ; Glycoproteins* ; Granulocytes* ; Humans ; Infant, Newborn ; Mass Screening ; Neutropenia ; Polymorphism, Restriction Fragment Length ; Pregnant Women ; Serotyping ; Thrombocytopenia ; Thrombocytopenia, Neonatal Alloimmune

BACKGROUND: Umbilical cord blood (UCB) has been increasingly utilized for reconstituting hematopoiesis in a variety of congenital disorders and hematologic malignancies. In this report, we evaluated the maximum collection volume, the efficacy of red cell depletion, cell viability and phenotypic analysis before and after cryopreservation. METHODS: Forty units of UCB (17 from NSVD and 23 from cesarean section) were collected into blood donation bag with ACD-A. Red cells were depleted using 3% gelatin sedimentation or buffy coat separation. UCB units were cultured in methylcellulose media, and phenotypic analyses were performed with monoclonal antibodies for CD34, HLA-DR, CD38, CD13, CD33, c-kit, CD45/CD3, CD45/CD19, CD3/CD4, CD3/CD8. RESULTS: The mean volume of one unit of UCB was 109.2 +/- 32.5 mL and one unit contained 1.20 +/- 0.51x19(9) nucleated cells. Cell counts after red cell depletion by 3% gelatin sedimentation or buffy coat separation revealed recovery rates of 77.5 +/- 14.9% and 64.5 +/- 7.4%, respectively. Cell viabilities and the number of colony forming units - granulocyte and monocyte from fresh and cryopreserved UCB were were 98.1 +/- 1.6%, 88.7 +/- 4.8%, and 6.48 +/- 2.14x10(5), 7.35 +/- 0.65x10(5). The mean percentage of CD34+ cells was 1.02 +/- 1.6% and those of CD34+/HLA-DR+, CD34+/CD38+, CD34+/CD13+, CD34+/CD33+, CD34+/c-kit+ cells were 68.6%, 58.0%, 5.6%, 46.8%, and 29.8%, respectively. Lymphocyte subsets were composed of CD45+/CD3+ (59.0%), CD45+/CD19+ (13.8%), CD3+/CD4+ (42.7%), and CD3+/CD8+ cells (17.1%). There was no significant difference in phenotypic characteristics between fresh and cryopreserved UCB. CONCLUSION: We established the protocols for UCB collection, red cell depletion, cryopreservation, culture and phenotypic analyses. These results would be very useful for future UCB transplantation and preservation/storage.
Antibodies, Monoclonal ; Blood Donors ; Cell Count ; Cell Survival ; Congenital, Hereditary, and Neonatal Diseases and Abnormalities ; Cryopreservation* ; Fetal Blood* ; Gelatin ; Granulocytes ; Hematologic Neoplasms ; Hematopoiesis ; HLA-DR Antigens ; Humans ; Lymphocyte Subsets ; Methylcellulose ; Monocytes ; Stem Cells ; Umbilical Cord*

BACKGROUND: Investigation of the factors affecting blood donation practice is essential to develop the ways of making blood donation campaign, as well as efficient and facilitating blood donation practice. A few studies has been made concerning the factors affecting blood donation in Korea. METHODS: 637 participants were examined using self-administered questionnaires including demographic variables, experience, knowledge and attitude for blood donation and others. RESULTS: 51.5% of participants had experienced the blood donation. Students who donated blood in high school days showed tendency to donate blood more than those who didn t donate blood in high school days. Students who had more knowledge and desirable attitude about blood donation experienced more blood donations. CONCLUSION: It is necessary for adolescents to take the opportunity of blood donation. It is important to clarify factors affecting blood donation practice and to encourage the public education and campaign which enable public has accurate knowledge and positive attitude about blood donation.
Adolescent ; Blood Donors* ; Education ; Humans ; Korea ; Surveys and Questionnaires

BACKGROUND: Screening of donor blood for malaria has not been activated in Korea yet in spite of the recent break out of tertian malaria. The microscopical and immunological methods, PCR in current use for malaria diagnosis in blood donations. We studied the positive rate of malaria antibody and antigen in army donation blood and samples of risky area (Daesung dong) as a screening method. METHODS: In this study, samples of Daesung dong residing people(n=77) and army donated blood donors(n=44) were screened for malarial antibody by Indirect Fluorescence Antibody (IFA) test and malarial antigen by Polymerase chain reaction(PCR) technique and microscopic exam of Wright Giemsa stain. RESULTS: A total of 6.7% blood donors showed positive result of malaria antibody by IFA test respectively, and 1.7% donors showed the presence of antigen both technique by the PCR and conventional microscopy. CONCLUSIONS: Army donated blood and samples of risky area had a high positive rate of malaria antibody and antigen, there need attention and effort of prevention of tranfusion transmitted malaria in donated blood from risky area.
Azure Stains ; Blood Donors ; Diagnosis ; Fluorescence ; Humans ; Korea ; Malaria* ; Mass Screening ; Microscopy ; Polymerase Chain Reaction ; Tissue Donors

BACKGROUND: Presence of fetal D positive RBCs in maternal peripheral blood of D negative pregnants induce Rh alloimmunization. Early detection of fetal D positive RBCs would eliminate the risk to an D positive fetus in pregnancy. We demonstrate and evaluate relibility of a sensitive polymerase chain reaction(PCR)-based assay for the RHD fetus gene detection from maternal peripheral blood samples. METHODS: The amplification of RHD gene and RHCcEe gene site were done in peripheral blood sample(n=17) of nine D negative pregnants by polymerase chain reaction and evaluated the sensitivity of PCR genotyping in serially diluted D positive sample. If RHD amplicated band(189 bp) were found with RHCcEe band(136 bp) in peripheral blood of D negative pregnants, the results were interpreted as positive for fetomaternal hemorrhage detection. RESULTS: Gestational age distribution of samples were from 7 to 39 weeks include postpartum one day. The RHD typing by PCR showed good sensitivity to detect up to 0.05% fetomaternal hemorrhage. Two cases were positive by PCR among 17 samples and their gestaional age was 22 and 27 weeks. CONCLUSIONS: RHD PCR showed good sensitivity for fetomaternal hemorrhage detection and prophylaxis of Rho(D) alloimmunization should be carefully done by this results because fetomaternal hemorrhage were detected in early gestational age by PCR.
Female ; Fetomaternal Transfusion ; Fetus ; Gestational Age ; Polymerase Chain Reaction* ; Postpartum Period ; Pregnancy

BACKGROUND: Preoperative autologous blood donation is utilized to reduce the risk of transfusion associated infections and transfusion reactions using homologous blood. We analyzed our experiences of autologous blood donations in Kyung Hee University Hospital. METHODS: From Jan. 1996 to Feb. 1997, we analyzed 213 patients who visited the blood donation room for autologous blood donation. RESULTS: Among 213 patients, 32 patients(15%) were rejected because of unfittedness with donor criteria. The majority of donor were orthopedic patients(95.4%). 14 patients(8%) could not finish the donation during the blood collection period because of donor reaction. The major cause was anemia(6 cases, 43%). During the donation period, mean of hemoglobin level was decreased by 1.3g/dL. Less than 1.5g/dL of hemoglobin reduction was observed in 117 patients(64.6%). In the use of donated blood, 43% used only autologous blood, 40.5% used autologous blood and additional homologous blood, and 16.5% were diacarded. The seropositive rates of HBsAg, anti-HCV, anti-HIV, and VDRL were 7%, 0.6%, 0%, and 0.6%, respectively. Simultaneously seropositive rate of HBsAg & anti-HCV was 0.6%. CONCLUSION: Autologous blood donation can be a good transfusion practice for elective surgery. Physician education programs are needed to avoid necessary homologous blood and unnececessary autologous blood donation.
Blood Donors* ; Blood Group Incompatibility ; Education ; Hepatitis B Surface Antigens ; Humans ; Orthopedics ; Tissue Donors