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Blood grouping
 

Blood grouping

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AGA KHAN UNIVERSITY HOSPITAL

AGA KHAN UNIVERSITY HOSPITAL

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    Blood grouping Blood grouping Presentation Transcript

    • BloodGrouping
    • History - Karl Landsteiner  Discovered the ABO Blood Group System in 1901  He and his five co-workers began mixing each others red cells and serum together and inadvertently performed the first forward and reverse ABO groupings
    • Landsteiners Rule  If an antigen (Ag) is present on a patients red blood cells the corresponding antibody (Ab) will NOT be present in the patients plasma, under ‘normal conditions’.
    • MajorABO BloodGroup ABO Group Antigen Present Antigen Missing Antibody Present A A B Anti-B B B A Anti-A O None A and B Anti-A&B AB A and B None None
    • ABO Basics  Blood group antigens are actually sugars attached to the red blood cell.  Antigens are “built” onto the red cell.  Individuals inherit a gene which codes for specific sugar(s) to be added to the red cell.  The type of sugar added determines the blood group
    • Principle of blood grouping There are two principles 1-almost all normal healthy individuals above 3-6 months of age have “ naturally occurring Abs” to the ABO Ags that they lack These Abs termed naturally occurring because they were thought to arise without antigenic stimulation
    • Principle of blood grouping 2- These “naturally occurring” Abs are mostly IgM class. That means that, they are Abs capable of agglutinating saline/ low protein suspended red cell without enhancement and may activate complement cascade.
    • ABO and H Antigen Genetics  Genes at three separate loci control the occurrence and location of ABO antigens  The presence or absence of the A, B, and H antigens is controlled by the H and ABO genes
    • Location  The presence or absence of the ABH antigens on the red blood cell membrane is controlled by the H gene  The presence or absence of the ABH antigens in secretions is indirectly controlled by the Se gene
    • HAntigen  The H gene codes for an enzyme that adds the sugar fucose to the terminal sugar of a precursor substance (PS)  The precursor substance (proteins and lipids) is formed on an oligosaccharide chain (the basic structure)
    • RBC Precursor Structure Glucose Galactose N-acetylglucosamine Galactose Precursor Substance (stays the same) RBC
    • Formation of the H antigen Glucose Galactose N-acetylglucosamine Galactose Precursor Substance (stays the same) RBC H antigen Fucose
    • H antigen  The H antigen is the foundation upon which A and B antigens are built  A and B genes code for enzymes that add an immunodominant sugar to the H antigen  Immunodominant sugars are present at the terminal ends of the chains and confer the ABO antigen specificity
    • A and B Antigen  The “A” gene codes for an enzyme (transferase) that adds N-acetylgalactosamine to the terminal sugar of the H antigen  N-acetylgalactosaminyltransferase  The “B” gene codes for an enzyme that adds D- galactose to the terminal sugar of the H antigen  D-galactosyltransferase
    • Formation of theA antigen Glucose Galactose N-acetylglucosamine Galactose RBC Fucose N-acetylgalactosamine A antigen
    • Formation of the B antigen Glucose Galactose N-acetylglucosamine Galactose RBC Fucose Galactose B antigen
    • H antigen  Certain blood types possess more H antigen than others: O>A2>B>A2B>A1>A1B
    • Why do Group O individuals have more H antigen than the other groups? Group O individuals have no A or B genes to convert the H antigen to A or B antigens….that means more H antigen sites
    • Group O Group A Many H antigen sites Fewer H antigen sites A A A AA Most of the H antigen sites in a Group A individual have been converted to the A antigen
    • Genetics  The H antigen is found on the RBC when you have the Hh or HH genotype, but NOT from the hh genotype  The A antigen is found on the RBC when you have the Hh, HH, and A/A, A/O, orA/B genotypes  The B antigen is found on the RBC when you have the Hh, HH, and B/B, B/O, orA/B genotypes
    • Bombay Phenotype (Oh)  Inheritance of hh  The h gene is an amorph and results in little or no production of L-fucosyltransferase  Originally found in Bombay  Very rare (130 worldwide)
    • Bombay Phenotype (Oh)  The hh causes NO H antigen to be produced  Results in RBCs with no H, A, or B antigen (patient types as O)  Bombay RBCs are NOT agglutinated with anti-A, anti- B, or anti-H (no antigens present)  Bombay serum has strong anti-A, anti-B and anti- H, agglutinating ALL ABO blood groups  What bloodABO blood group would you use to transfuse this patient?? Another Bombay Group O RBCs cannot be given because they still have the H antigen You have to transfuse the patient with blood that contains NO H antigen
    • ABO antibodies  GroupA serum contains anti-B  Group B serum contains anti-A  GroupAB serum contains no antibodies  GroupO serum contains anti-A, anti-B, and anti-A,B
    • ABO antibodies  IgM is the predominant antibody in GroupA and Group B individuals  Anti-A  Anti-B  IgG (with some IgM) is the predominant antibody in GroupO individuals  Anti-A,B (with some anti-A and anti-B)
    • ABO antibodies  Reactions phase: Room temperature  Complement can be activated with ABO antibodies (mostly IgM, some IgG)  High titer: react strongly (4+)  Usually present within the first 3-6 months of life  Stable by ages 5-6 years  Decline in older age  Newborns may passively acquire maternal antibodies (IgG crosses placenta)  Reverse grouping (with serum) should not be performed on newborns or cord blood
    • ABO routine testing  Several methods for testing the ABO group of an individual exist.The most common method is:  Serology: This is a direct detection of the ABO antigens. It is the main method used in blood transfusion centres and hospital blood banks.  This form of testing involves two components: a)Antibodies that are specific at detecting a particular ABO antigen on RBCs.  b) Cells that are of a known ABO group that are agglutinated by the naturally occurring antibodies in the person's serum.
    • ABO ROUTINE TESTING DIRECT OR FORWARD GROUPING Test for antigens • Patient’s cells containing unknown antigens tested with known antisera • Antisera manufactured from human sera Aantisera used: Antisera Color Source Anti-A Blue Group B donor Anti-B Yellow Group A donor Anti-A,B Red Group O donor
    • Forward Grouping  Reaction of patient red blood cells tested with Reagent anti-A and anti-B antisera  Slide: 20-40% RBC suspension + anti-serum  Tube (12x75mm): 2-5% RBC suspension + anti-serum (centrifuge before read)
    • Forward Grouping Reaction Patterns for ABO Groups Blood group Agglutination with Anti-A Agglutination with Anti-B A + - B - + AB + + O - -
    • Reverse grouping • serum is combined with cells having known Ag content in a 2:1 ratio • uses commercially prepared reagents containing saline-suspended A1 and B cells
    • Reverse grouping Reaction Patterns for ABO Groups Blood Group Agglutination with A cells Agglutination with B cells A - + B + - AB - - O + +
    •  Grading of Agglutination: Negative (0) No clumps or aggregates Weak (+/-) Tiny clumps or aggregates barely visible macroscopically or to the naked eye 1+ Few small aggregates visible macroscopically 2+ Medium-sized aggregates 3+ Several large aggregates 4+ One solid aggregate
    • ABO blood group (forward blood grouping) Patient Red CellsTested With InterpretationAnti-BAnti-APatient 001 04+2 4+03 4+4+4
    • ABO blood group (forward blood grouping) Patient Red CellsTested With InterpretationAnti-BAnti-APatient O001 A04+2 B4+03 AB4+4+4
    • Reverse Grouping (Confirmatory grouping Patient SERUMTestedWith InterpretationB CellsA1 CellsPatient 4+4+1 4+02 04+3 004
    • Reverse Grouping (Confirmatory grouping Patient SERUMTestedWith InterpretationB CellsA1 CellsPatient O4+4+1 A4+02 B04+3 AB004
    • Forward & reverse ABO blood grouping Reaction of CellsTested With Reaction of SerumTested Against ABO Group Anti-A Anti-B A1 Cells B Cells 1 0 0 + + O 2 + 0 0 + A 3 0 + + 0 B 4 + + 0 0 AB
    • Forward & reverse ABO blood grouping Reaction of CellsTested With Reaction of SerumTested Against ABO Group Anti-A Anti-B A1 Cells B Cells 1 0 0 + + 2 + 0 0 + 3 0 + + 0 4 + + 0 0
    • ID card system  This ID-Card contains a mixture of human polyclonal and monoclonal anti-A, human polyclonal anti-B and human polyclonal anti-D antibodies.  The microtube ctl is the negative control.Two microtubes with neutral gel serve for reverse grouping with A1 and B cells.
    •  Fully-automatic walk-away for ID-Cards  Stand alone instrument  Continuous sample loading  Continuous reagent loading  Priority samples (STAT)  Reagent stock  High throughput
    •  Optimized for small blood volumes  Dispense verification  Easy-to-use  Full test menu  Wi-Fi  Touchscreen 17"  Host connectivity  Internal & external validation  Capacity 180 samples 240 ID-Cards 28 reagent vials
    • Thank you….