Agglutination Tests ; methods ; Brucellosis ; blood ; diagnosis ; Coombs Test ; methods ; Humans ; Sensitivity and Specificity

BACKGROUND: The results of a particular test can be affected by the techniques used for testing. However, limited data is available on the testing techniques used in blood bank laboratories in Korea. The aim of this study was to evaluate the various testing techniques used in blood bank laboratories using the data obtained during the past six years from the Korean external quality assessment (KEQA) of blood bank laboratories. METHODS: Data was collected from all KEQA respondents via the KEQA website on the testing techniques used in blood bank laboratories from 2008 to 2013. The survey included questions on ABO grouping, D typing, crossmatching tests, direct antiglobulin tests (DAT), antibody (Ab) screening, and Ab identification (ID) tests. RESULTS: Based on the data obtained from 2008 to 2013 (ABO grouping data obtained from 2011 to 2013), the most frequently used techniques are as follows: slide agglutination (60.7% and 60.8%) for ABO cell typing; tube agglutination (78.2% and 81.2%) for ABO serum typing; slide agglutination (50% and 54.6%) for D typing; tube agglutination (91.9% and 83.8%) for crossmatching tests; tube agglutination (63.6% and 52.8%) for DAT; column agglutination technique (CAT; 74.5% and 89.4%) for Ab screen; and CAT (83.9% and 94.2%) for Ab ID. CONCLUSIONS: The findings reveal a steady increase in the use of CAT from 2008 to 2013 for crossmatching tests, DAT, Ab screen, and Ab ID and a decreasing use of the tube agglutination technique for the past six years. Since the slide agglutination technique accounted for a significant percentage of the tests conducted, further education is warranted on the improvement in the techniques used for ABO and D typing.
Agglutination ; Agglutination Tests ; Animals ; Blood Banks* ; Cats ; Clinical Laboratory Techniques ; Coombs Test ; Surveys and Questionnaires ; Education ; Korea ; Mass Screening

BACKGROUND: Blood loss due to laboratory phlebotomy among neonates is correlated with anemia as well as transfusion. In this study, microcolumn agglutination cards for performing ABO & RhD blood typing and direct antiglobulin tests in neonates were evaluated and compared with other established systems. Also, the blood group antibody production rates according to the age were calculated to determine the upper age limit for the new method. METHODS: Eighty subjects were tested by using the DianaGel Neonatal cards (Diagnostic Grifols, Barcelona, Spain), and the results were compared with those of the slide methods for ABO and RhD blood typing, and the DiaMed-ID DC-Screening I test (DiaMed, Morat, Switzerland) for direct antiglobulin tests. A total of 546 subjects who were under 12 months old were tested for the ABO back-typing, and 58 subjects with the AB blood type were excluded. RESULTS: The results of the DianaGel Neonatal card were in agreement with those of the conventional methods for all the subjects. Only one subject showed a discrepant result for the DAT between the DianaGel and DiaMed methods. Blood group antibodies were detected in 29 out of 169 (17.2%) one-day-old neonates, in eight out of 34 (23.5%) infants between one and three months of age and in 81 out of 96 (84.4%) infants between six and twelve months of age. CONCLUSION: The DianaGel Neonatal card showed at least equivalent performance as compared to that of the conventional methods, and it showed advantages for a low blood volume requirement and stronger agglutination grades. The DianaGel card is a suitable alternative for blood typing and DAT in infants under the age of 3 months and who do not necessarily need back-typing of the blood groups due to the low production rate of antibodies.
Agglutination ; Anemia ; Antibodies ; Antibody Formation ; Blood Group Antigens ; Blood Grouping and Crossmatching ; Blood Volume ; Coombs Test ; Humans ; Infant ; Infant, Newborn ; Phlebotomy

No absract available.
Desensitization, Immunologic*

No abstract available.
Desensitization, Immunologic

The purpose of this study was to determine whether the fully automated ORTHO AutoVue Innova system, which based on the microcolumn glass sphere technology, is accurate enough to meet immunohematology testing needs at blood banks. 16 IgM anti-C, anti-c, anti-D, anti-E and anti-e dilution series were tested respectively, with corresponding antigen positive red blood cell solutions, by ORTHO AutoVue Innova system and saline medium test. 16 IgG anti-D dilution series were tested respectively with RhD positive red blood cell solutions by ORTHO AutoVue Innova system, polybrene test and antiglobulin test. The accuracies of microcolumn glass sphere technology were analysed, by comparing to the reference assays. The results showed that the sensitivities of the ORTHO AutoVue Innova tests were 1:69.8, 1:33.4, 1:1448.1, 1:139.6 and 1:32.0 for IgM anti-C, anti-c, anti-D, anti-E and anti-e respectively; the corresponding value of saline medium tests were 1:16.7, 1:16.6, 1:430.5, 1:34.9 and 1:9.9. There were statistically significant differences between the groups of each tests (t values were 14.38, 5.48, 10.25, 12.65 and 9.59 for IgM anti-C, anti-c, anti-D, anti-E and anti-e respectively, p < 0.05). For IgG anti-D, the sensitivities of the ORTHO AutoVue Innova test, polybrene test and antiglobulin test were 1:980.6, 1:181.0 and 1:304.4 respectively. There was statistically significant difference among the 3 groups (F = 51.15, p < 0.01). It is concluded the use of ORTHO AutoVue Innova system for blood group compatibility test can obtain more accurate results than traditional tube tests, it is reliable and safe for routine tests performed in immunohematology laboratories.
Blood Grouping and Crossmatching ; methods ; Coombs Test ; methods ; Humans ; Isoantibodies ; blood ; Materials Testing ; Rho(D) Immune Globulin ; Sensitivity and Specificity

BACKGROUND: Blood typing is an essential test for transfusion. Generally, blood typing is performed using a slide test, tube test or microcolumn agglutination test. The aims of this study were to develop a new blood typing kit using micromachining, microfluidics and microseparation methods, and to evaluate the clinical usefulness of the new blood typing kit. METHODS: We designed and manufactured a blood typing microchip using polydimethylsiloxane (PDMS), which contained a microchannel (25~200 micrometer). The blood sample and antisera to be tested were dropped on the microwell for movement and mixing by capillary action. Once agglutination occurred, the microchannel acts as a filter and the blood type was determined by observation by the naked eye. To evaluate the newtyping kit, we tested sensitivity using artificially diluted blood and compared the results of the new typing method with the slide and tube methods using 70 samples. RESULTS: The new blood typing kit could differentiate a +4~+2 agglutination reaction, but could not detect a +1 agglutination reaction as observed by the naked eye. Among 70 samples, the results of ABO and Rh typing by the new typing method (n=66, > or = +2 agglutination reaction by the column agglutination method) were in accord with the results of the tube and slide methods, but couldnot detect agglutination in all 4 clinical samples, below a +1 agglutination reaction. CONCLUSION: The new blood typing kit is inadequate for routine use in the clinical laboratory due to low sensitivity, but with further improvement, it can be used economically, conveniently and objectively for blood typing without any special equipment. Moreover, the microfludics and separation method may be broadly applicable in other tests using the hemagglutination method.
Agglutination ; Agglutination Tests ; Blood Grouping and Crossmatching* ; Capillary Action ; Hemagglutination ; Immune Sera ; Microfluidics* ; Microtechnology

BACKGROUND: The weak D is characterized serologically by a weak or negative agglutination reaction with polyclonal anti-D in an immediate-spin test and agglutination is enhanced in the indirect antiglobulin test. Weak D has a lower number of D antigen or weaker antigen density than are normal D positive red cells. Here we studied the cause of weak D antigenicity at genetic level and compared to that of normal D RBCs. METHODS: The amplification of RHD gene and RHCcEe gene site was done in normal D(n=20), weak D(n=8), D negative group(n=20) by polymerase chain reaction and by based on D typing in these individuals compared to that of serologic D typing. In addition, to detect RHD gene mutation and nucleotide sequence difference of weak D group compared to normal D RBCs, single stranded conformational polymorphism PCR was simultaneuosly perfomed in two group by RHD amplified product(189 bp). We analysis the correlation RHD genotyping and serological phenotyping, and also analysis the difference of nucleotide sequence between two group in genetic level. RESULTS: The RHD genotyping was completely matched normal D(n=20), D negative group(n=20) but weak D group(n=8) showed same genotype of normal D RBCs. In single stranded conformational polymorphism PCR, weak D phenotypes does not show any abnormalities at the genomic level when compared to the RHD gene in normal D phenotypes. CONCLUSIONS: RHD PCR showed good correlation with conventional serologic test but weak D genotype was same as that of normal D RBCs. The weaker immunogenicity of weak D is not explained by genomic DNA difference itself.
Agglutination ; Base Sequence ; Coombs Test ; DNA ; Genotype ; Phenotype ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Serologic Tests

BACKGROUND: Irregular antibodies are antibodies that are not regularly present in the serum of particular blood groups and its presence results in many problems including HDN (hemolytic disease of newborn) in transfusion medicine. Column agglutination test was recently introduced and has been widely used for advantages of standardized working procedures, standard reactions, stable reactions for hours and Coombs test without washing steps. We tested irregular antibodies in cord blood by column agglutination test and investigated its incidence and relation with HDN. METHODS: We tested the cord blood collected during delivery from 200 pregnant women. Column agglutination test was done on DiaMed ID MicroTyping System (DiaMed, Switzerland) and both LISS/Coombs and NaCl/Enzyme ID-cards were used. The antibody screening test was done first and antibody identification test was done to positive cases in same way. The cell typing and Rh phenotyping for cord blood of positive cases were also done. RESULTS: 2 cases of 200 samples (1%) were positive in the antibody screening test and each was identified as anti-D and anti-E antibody. CONCLUSIONS: Irregular antibody screening in cord blood by column agglutination test is thought to be helpful in early diagnosis and treatment of HDN.
Agglutination Tests* ; Agglutination* ; Antibodies ; Blood Group Antigens ; Coombs Test ; Early Diagnosis ; Erythroblastosis, Fetal ; Female ; Fetal Blood* ; Humans ; Incidence ; Infant, Newborn ; Mass Screening* ; Pregnant Women ; Transfusion Medicine

Allergen immunotherapy (AIT) and diagnostic tests are based on well qualified allergen extracts, which are derived from biologic organisms. The allergenicity of the extracts is markedly affected by the climate, soil, year of production, storage methods, and manufacturing processes. Thus, standardization is a crucial process to guarantee the clinical efficacy and safety of the treatment and diagnostic reagents in allergic diseases. There are 2 different standardization processes, one is In vivo and the other is in vitro standardization. In vivo standardization is done by skin prick or intradermal tests. For in vitro standardization, measurements of weight/volume and protein nitrogen units have been widely used since the early period of AIT. In the 1970s, immunological methods such as radial immunodiffusion, enzyme-linked immunosorbent assay (ELISA) inhibition test and basophil activation test were developed. Allergen potency measured by ELISA inhibition test reflects the potency measured by skin tests and has been widely used for quality control of batch-to-batch variation. Recently, standardizations focused on the major allergen content of extracts have developed. Standardization for major allergens requires reliable reference materials (RMs) made of recombinant allergens and 2-site ELISA kits. However, only a few reliable RM and 2-site ELISA kits are available. For the standardization process, allergen RMs are essential. The Center for Biologics Evaluation and Research of the U.S. Food and Drug Administration provides 19 allergen RMs, and our research team also proved 9 RMs which are important in Korea. In conclusion, allergen standardization is an essential process for the development of reliable treatment and diagnostic reagents, and allergy specialist should be familiar with the concept of allergen standardization.
Allergens ; Basophils ; Biological Products ; Climate ; Desensitization, Immunologic ; Diagnostic Tests, Routine ; Enzyme-Linked Immunosorbent Assay ; Hypersensitivity ; Immunodiffusion ; In Vitro Techniques ; Indicators and Reagents ; Intradermal Tests ; Korea ; Nitrogen ; Quality Control ; Skin ; Skin Tests ; Soil ; Specialization ; Treatment Outcome ; United States Food and Drug Administration