1st Assessment: Blood bank Reagents and Methods

What are the sources of Ag & Ab in ABO/Rh typing?

• Source of antigens:

  • Patient's RBCs (forward grouping/typing);

  • Reagent RBCs (reverse grouping)

• Source of antibodies:

  • Reagent anti-A, anti-B, and anti-D (forward grouping/typing);

  • patient's serum (reverse grouping)

A test that detects specific antbodies to RBC antigens

Antibody screen

A test that identifes Abs to RC Ags

Antibody identification

What are the sources of Ag & Ab in Antibody Screening?

• Source of antigens:

  • Reagent antibody screening cells

• Source of antibodies:

  • Patient's serum

What are the sources of Ag & Ab in Antibody Identification?

• Source of antigens:

  • Reagent antibody panel cells

• Source of antibodies:

  • Patient's serum

A test that detects A, B, and D antigen

ABO/Rh typing

What are the sources of Ag & Ab in Crossmatching?

• Source of antigens: Donor cells

• Source of antibodies: Recipient's serum

A test that determines the compatibility of donor RBCs with recipient’s blood

Crossmatch

Possess known antigens and are treated to prolong their life span

Reagent RBCs

Contain known antibodies against specific RBC antigens

Antisera

Contain poly- or monospecific antibodies against human antibodies

Antiglobulin reagents

• Blood bank reagents are licensed by the ___ and ___

• ____ specifies the potency and the specificity of reagnts before production

• Center for Biologies Evaluation and Research of the Food and Drug Administration (FDA)

• FDA

What are the types of Reagent Antisera?

1. Polyclonal

2. Monoclonal

3. Blended Monoclonal

4. ABO antisera

5. D-Typing

Antibodies are IgM and react best at ____ or ____

room temperature or 4°C.

A reagent that is often used in direct antiglobulin tests (DAT)

Polyspecific

Types of Antiglobulin Reagents

1. Polyspecific

2. Monoclonal

It is used as a control to ensure antiglobulin reagent reactivity in each negative antiglobulin test tube

-IgG-coated control cells (Check cells):

Types of Reagent RBC’s :

1. IgG coated control cells

2. A1 and B-cells

3. Antibody ponel cellls

This technique uses dextran acrylamide gel combined with reagents or diluent. Anti-IgG cards are used for DATs and lATs.

Gel technology

• The traditional tube method is adapted to the microtiter plate, in which smaller volumes of serum and cells are used, and it is read on an automated photometric instrument.

• The cell buttons are resuspended by tapping the sides of the plate.

Microplate methods

• RBC screening cells are bound to surface of microtiter plates.

• Negative

  • RBC button;

• Positive

  • RBCs (indicator cells) on sides and bottom of wells

Solid-phase adherence methods

Used to detect in vitro sensitization of RBCs.

Indirect antiglobul in test (IAT)

• In this procedure, reagent red cells are mixed with patient's serum, then incubated at 37°C to allow IgG antibodies to attach to the RBCs

• solution is then washed to remove unbound proteins.

• AHG is added to detect in vitro sensitization of RBCs

Indirect antiglobul in test (IAT)

Reagents added to the in vitro antiglobulin test to enhance antigen-antibody complex formation

Potentiator

increases antibody uptake of antigen

LISS

allows sensitized cells to come close together to form agglutination lattices

Bovine albumin (22% or 30%)

concentrates antibodies and creates a low-ionic solution to allow greater antibody uptake

Polyethylene glycol (PEG) additive

The proteolytic enzymes

1. Papain

2. Ficin

3. Bromelin

Remove certain structures from the RBC and enhance the access of antibodies to other less superficial structures on the RBC

Enzyme

What are the Potentiating media (AB enhacer)

1. LISS

2. Bovine Albumin

3. Polyethylene glycol

4. Proteolytic Enzyme