ABO DISCREPANCIES. Objectives: Upon completion of this CLS 422 unit, the Clinical Laboratory Science student will:
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1 ABO DISCREPANCIES 1. Recognize any discrepancies when given sets of reactions for ABO typing. 2. Identify possible reasons for unexpected reactions of the ABO system in: a. Cell grouping b. Serum grouping 3. Propose steps that could correct or confirm ABO discrepancies. CLS 422 Clinical Immunohematology I Objectives Page 1 of 14
2 THE ABO SYSTEM 1. Discuss the discovery of the ABO system. 2. Evaluate the reactions in both cell and serum typing of ABO groups, using conventional reagents. 3. Determine the ABO typing results using conventional reagents, for each of the following subgroups: a. A 1 b. A 2 4. Explain the inheritance of the A, B and H genes. 5. Name the specific transferases and terminal sugars for the A, B and H substances. 6. Predict all the possible genotypes of children, given the ABO phenotype of parents. 7. Assess the secretor gene s influence on A, B and H substances found in the secretions of persons of various ABO groups. 8. Discuss the importance of the ABO system as applied to transfusion medicine. 9. Evaluate the significance of A subgroups in the clinical situation. 10. State the sources and uses of the following: a. Absorbed anti-a 1 b. A 1 lectin c. H lectin 11. Evaluate the following characteristics of the ABO system: a. Class of immunoglobulin (antibody) b. Enzyme effect c. Complement activation 12. State the genotype of individuals with the Bombay phenotype. 13. State the comparative concentrations of H substance in each ABO phenotype. 14. Differentiate between Bombay anti-h and auto anti-h. CLS 422 Clinical Immunohematology I Objectives Page 2 of 14
3 ABSORPTION AND ELUTION 1. Define the following terms: a. Absorption b. Adsorption c. Elution d. Eluate 2. Evaluate situations in which each of the following may be indicated: a. Absorption studies b. Elution studies c. Absorption and elution studies 3. Compare the following techniques for preparing eluates: a. Heat/freeze b. Alteration of ph c. Chemical elution (chloroquine, ZZAP) 4. Explain technical factors associated with absorption and elution procedures. 5. Discuss the action of the following reagents used in absorption or elution studies: a. ZZAP b. Chloroquine diphosphate c. EDTA Glycine Acid (EGA) d. Rabbit Erythrocyte Stroma (RESt) e. Human Platelet Concentrate (HPC) f. 2-mercaptoethanol (2-ME) 6. Select the appropriate reagent listed in objective #5 for use, when given pertinent data. CLS 422 Clinical Immunohematology I Objectives Page 3 of 14
4 ANTIBODY DETECTION AND IDENTIFICATION 1. Explain the purpose of performing an antibody screen in each of the following groups: a. Prenatal patients b. Donors c. Blood recipients 2. Discuss the antigen characteristics important in the composition of screening cells. 3. Describe the phases of antibody detection. 4. Describe the types of antibodies that can be encountered in each of the phases of antibody detection. 5. Compare and contrast the following methods for performing an antibody screen: a. Tube b. Gel (microcolumn agglutination) c. Solid phase adherence 6. Explain the mechanism by which the four common enzymes used in Blood Bank are able to enhance or inhibit antigen-antibody reactions. 7. Define a panel of cells. 8. Recommend testing that can be performed to confirm the identification of antibody(ies). 9. Explain how the following factors aid in the interpretation of antibody panels: a. Cross-out technique b. Variation in strengths of reaction c. Phases of reaction d. Autocontrol e. Prewarming f. Saline replacement g. 95% confidence rule h. Cord cell results i. Neutralization j. Red cell antigen determinations k. Enzymes 10. Describe the steps that can be taken if a weakly reacting antibody is detected. 11. State the difference between an alloantibody and an autoantibody. CLS 422 Clinical Immunohematology I Objectives Page 4 of 14
5 12. Interpret a set of reactions to determine when each of the following situations is present: a. Cold agglutinins b. Warm agglutinins c. Abnormal protein d. Single alloantibody e. Multiple alloantibodies 13. Determine the percentage of donors compatible with any antibody when given an antigen frequency chart. 14. Define the following terms: a. Panagglutination b. Polyagglutination c. Albumin-agglutinating phenomenon d. Antibody to a preservative e. Rouleaux formation 15. Identify the antibody(ies) present when given panel results. 16. Select additional testing to aid in the identification of antibody(ies) when given panel results. CLS 422 Clinical Immunohematology I Objectives Page 5 of 14
6 THE ANTIGLOBULIN TEST 1. Differentiate between monospecific and polyspecific anti-human globulin reagent according to each of the following: a. Composition b. Use c. Preparation 2. Explain the principle of the antiglobulin reaction. 3. Evaluate the Direct Antiglobulin Test (DAT) in the following situations: a. Hemolytic disease of the fetus and newborn (HDFN) b. Hemolytic transfusion reactions (HTR) c. Hemolytic anemias d. Drugs e. Diseases 4. Evaluate the significance of a positive Direct Antiglobulin Test in the following: a. Antigen typing b. Weak D testing c. Autocontrol d. Antiglobulin crossmatch 5. Evaluate the Indirect Antiglobulin Test in the following situations: a. Antigen typing b. Compatibility testing c. Antibody detection d. Antibody identification 6. Differentiate between the indirect and direct antiglobulin procedures. 7. List at least four factors affecting the antigen-antibody reactions in the antiglobulin test. 8. Assess sources of error affecting the antiglobulin test. 9. Name the control system for the antiglobulin test. 10. Interpret both positive and negative check-cell results. CLS 422 Clinical Immunohematology I Objectives Page 6 of 14
7 BASIC IMMUNOLOGY 1. Define the term immunohematology. 2. Define the following terms: a. Antigen b. Antibody c. Agglutinin d. Hemolysin e. Complement f. Humoral response g. Cellular response h. Immune response i. Zeta potential j. Homozygous k. Heterozygous l. Dosage 3. Differentiate between phenotype and genotype. 4. Describe the role lymphocyte populations have in the humoral and cellular responses. 5. Differentiate between primary and secondary immune responses. 6. Describe the basic immunoglobulin structure. 7. Describe the structural and functional characteristics and the serum concentration of the five immunoglobulin classes. 8. Compare IgG and IgM based on the following characteristics: a. Phase of reaction b. Immune versus naturally occurring c. Placental transfer d. Complement activation 9. Apply Landsteiner s Law to antibody formation. 10. Explain four factors that influence antigen-antibody reactions. 11. Describe the various methods used for in vitro detection of antigen-antibody reactions. CLS 422 Clinical Immunohematology I Objectives Page 7 of 14
8 BLOOD COMPONENTS AND DERIVATIVES 1. Outline the procedure for preparation of each of the following components, which may be obtained from a unit of whole blood or apherisis, if applicable: a. Packed red blood cells (RBCs) b. Platelet concentrate c. Fresh frozen plasma (FFP) d. Cryoprecipitate e. Washed RBCs and platelets f. Leukoreduced packed RBCs and platelets g. Frozen RBCs 2. Compare four anticoagulants used for blood preservation. 3. Describe the storage lesion of donor blood. 4. State the expected survival time of transfused RBCs and platelets. 5. Evaluate the indications and contra-indications for transfusion of each component listed in Objective #1. 6. Describe the temperature and length of storage and transportation for each component listed in Objective #1. 7. State the important coagulation factors found in each of the following: a. Fresh frozen plasma b. Cryoprecipitate 8. Discuss the quality control (coagulation factors, number of platelets, hematocrit, etc.) specific for each component listed in Objective #1. 9. Distinguish between the following kinds of pheresis, including applications for each: a. Plateletpheresis b. Leukopheresis c. Plasmapheresis 10. Discuss the use of HLA-matched plateletpheresis. 11. Discuss reasons for requiring the following blood component attributes: a. CMV-negative b. Irradiated c. Leukoreduced d. Hemoglobin S negative e. Fresh f. Phenotypically matched CLS 422 Clinical Immunohematology I Objectives Page 8 of 14
9 12. Explain criteria for donor blood inspection and quarantine procedures, as outlined in AABB standards. 13. Explain the benefit of giving the following derivatives, as opposed to blood components: a. Factor VIII concentrate b. Factor IX concentrate c. Albumin d. Gamma globulin 14. Discuss the various synthetic RBC/hemoglobin substitutes. 15. Select the appropriate component and/or derivative for a patient, when given the diagnosis and laboratory data on that patient. CLS 422 Clinical Immunohematology I Objectives Page 9 of 14
10 HEMOLYTIC DISEASE OF THE FETUS AND NEWBORN 1. Describe four criteria necessary for the occurrence of hemolytic disease of the fetus and newborn (HDFN). 2. Recognize possible consequences of HDFN. 3. Compare ABO HDFN and Rh HDFN. 4. Name antibodies frequently responsible for HDFN in descending order of severity. 5. Discuss the importance of each of the following prenatal screening tests: a. ABO grouping b. Rh typing c. Antibody screening d. Antibody titers e. Amniocentesis, including the use of the Liley Graph f. Ultrasound 6. Discuss the theory of action of Rh immune globulin (RhIg). 7. Discuss the effect of Rh immune globulin (RhIg) on each of the following: a. Antibody screen b. Antibody panel c. Antibody titer 8. Analyze the following criteria for possible RhIg administration: a. Mother s Rh type b. Newborn s Rh type c. Mother s anti-d status d. Time frame (prenatal and postnatal) 9. Assess the results of laboratory tests frequently performed on cord blood to diagnose and evaluate HDFN. 10. Explain the principle of the screening test to detect fetal maternal hemorrhage in the Rhnegative mother. 11. Discuss the principle and the use of the Kleihauer-Betke test. 12. Calculate the number of RhIg vials to be administered, when given the number of fetal cells in the mother s circulation. 13. Analyze the effect of repeated intrauterine transfusions on newborn testing. CLS 422 Clinical Immunohematology I Objectives Page 10 of 14
11 14. Explain the benefits of the following procedures used to treat neonates with HDFN: a. Intrauterine transfusion b. Induction of labor c. Phototherapy d. Neonatal transfusion e. Exchange transfusion 15. Select the appropriate specimen to use for compatibility testing in exchange transfusions. 16. Select appropriate blood for exchange transfusion. 17. Correlate laboratory data with clinical findings to determine the presence of HDFN. CLS 422 Clinical Immunohematology I Objectives Page 11 of 14
12 OTHER BLOOD GROUP SYSTEMS 1. List the antigens and corresponding antibodies of the following blood group systems: a. Lewis (Le) b. Kell (K, k, Js, Kp) c. Duffy (Fy) d. Kidd (Jk) e. MNSs f. P g. Ii 2. Discuss the discovery of the Kell blood group system. 3. State the optimal conditions of reactions for the antibodies of each system listed in objective #1. 4. Evaluate the following characteristics of each system listed in objective #1: a. Effect of enzymes b. Antigen variability c. Disease association d. Complement activation e. Dosage 5. Explain the inheritance of each blood group system listed in objective #1. 6. Assess the clinical significance of each blood group system listed in objective #1 in regards to hemolytic transfusion reactions(htr) and hemolytic disease of the fetus and newborn (HDFN). 7. Define high and low prevalence antigens. 8. Recognize examples of high and low prevalence antigens. 9. Define HTLA antibody. 10. Explain the clinical significance of HTLA antibodies. CLS 422 Clinical Immunohematology I Objectives Page 12 of 14
13 PRETRANSFUSION COMPATIBILITY TESTING 1. Explain the purpose of pretransfusion compatibility testing and include specifically what it will and will not insure. 2. Evaluate the following procedures that are required before components are issued for transfusion according to AABB Standards: a. Specimen acceptability b. ABO and Rh typing of recipient s blood c. Red cell antibody detection tests using recipient s serum or plasma d. Comparison of current findings on recipient s sample with records of previous results e. Tests on donor blood f. Selection of components of ABO and Rh types appropriate for the recipient g. Performance of a serologic or computer crossmatch h. Labeling of components with the recipient s identifying information i. Emergency release protocol 3. Prioritize available units for transfusion. 4. Interpret the significance of the phase in which an incompatibility occurs. 5. Select appropriate procedures to complete compatibility testing when confronted with each of the following situations: a. Single alloantibody b. Multiple alloantibodies c. Abnormal protein d. Positive direct antiglobulin test (DAT) on the recipient s red blood cells e. Positive DAT on the donor s red blood cells 6. State the requirements for retention of donor and recipient blood samples, according to AABB Standards. 7. Discuss AABB requirements for issuing blood components. 8. Discuss appropriate blood administration procedures, including acceptance criteria for returning a unit to inventory after issue. CLS 422 Clinical Immunohematology I Objectives Page 13 of 14
14 THE Rh SYSTEM 1. Compare the three theories of inheritance of the Rh antigens. 2. List the antigens and antibodies of the system using Wiener, Fisher-Race, and ISBT nomenclature. 3. Translate Rh haplotypes between Wiener, Fisher-Race and ISBT nomenclatures. 4. Determine all possible genotypes when given Rh phenotypes. 5. Calculate the most probable genotype when given Rh phenotypes. 6. Evaluate the following characteristics of the Rh system: a. Dosage b. Enzyme effected c. Complement activation d. Antigen variability e. Antigen structure 7. Evaluate each of the following as it relates to Rh typing sera and controls: a. Acceptable reagents b. Appropriate use 8. Propose steps that should be taken if a patient has a positive Rh control. 9. Differentiate the three theories of the weak D antigen. 10. Discuss situations when weak D testing would be appropriate. 11. Interpret the significance of results of the weak D procedure in each of the following circumstances: a. Positive direct antiglobulin test (DAT) b. Donor typing c. Patient typing 12. Discuss the clinical importance of the Rh system in transfusion. 13. Explain the clinical significance of the following compound and/or variant antigens in the Rh system: a. C w b. f c. G 14. Discuss the genetic mechanisms resulting in gene-deleted and Rh null genotypes. 15. Describe the impact of the Rh null genotype on the individual (i.e., Rh null disease). CLS 422 Clinical Immunohematology I Objectives Page 14 of 14
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