Lab 3 26

Blood Biochemistry BCH 471[Practical]
Lab (3) ABO Blood Grouping & Rh Groups

Objectives
1. To determine the blood group according to the ABO system.
2. To test for the availability of the Rh factor (D antigen).

💡 Remember !!
Blood Group Substances Antigen ☓ Antibody
 The differences in human blood are due to the presence or absence of certain protein molecules called antigens
and antibodies.
 The antigens are glycoproteins located on the surface of the red blood cells.
 The antibodies are proteins present in the plasma to attack foreign antigens, resulting in clumping (agglutination).
 ABO blood grouping consists of:

1. Two antigens (A & B) on the surface of the RBCs
2. Two antibodies in the plasma (anti-A & anti-B)

Antigen
Antibody
RBC
ABO Blood Type System
 The ABO blood type system is the major blood type classification system.
 The four blood types in the ABO system (A, B, AB, and O) refer to different versions of glycoproteins which
are present on the surface of RBCs.
Blood Types:
Surface Plasma
Blood Type
antigens antibodies
A-surface
Type A Anti-B
antigens
B-surface
Type B Anti-A
antigens
Type AB A and B antigens No antibodies
No surface
Type O Anti-A and anti–B
antigens
Importance of The ABO System
 Blood group antigens must be determined to secure a safe practice of blood transfusion.
 They are also useful in determining familial relationships in forensic medicine.

Genetics of Blood Types
 Individuals inherit a gene which codes for specific antigen(s) to be added to the red cell.
 You have two copies of this gene, one inherited from your mother and the other inherited from your father.
 There are 3 versions ‘alleles’ for blood type: A, B & O.
 Since we have 2 genes, there are 6 possible combinations.
 The A and B genes are dominant and the O gene is recessive.
Blood Antibody
Mother Genotypes
type Present
Father
A B O
A+A A Anti-B
A AA AB AO A+O A Anti-B
A+B AB None
B BA BB BO B+B B Anti-A
B+O B Anti-A
O+O O Anti A&B
O OA OB OO
Codominance
Is a condition in which the alleles of a gene pair in a heterozygote are fully expressed
thereby resulting in offspring with a phenotype that is neither dominant nor recessive.
Rhesus Blood Group
 First studied in rhesus monkeys.
 Is the second most significant blood group system in human transfusion.
 The D antigen (RhD) is the most important.
 If it is present on RBCs’ surface, the blood is RhD positive (~80% of the population), if not it's RhD negative.
Some people in group A will have it, and will therefore be classed as A+ (or A positive), while the ones that don't,
are A- (or A negative) and so it goes for groups B, AB and O.
Rh Blood Group Transfusion
 A person with Rh+ blood can receive blood from a person with Rh- blood without any problems.
 A person with Rh- blood can develop Rh antibodies in the blood plasma if he or she receives blood from a
person with Rh+ blood, whose Rh antigens can trigger the production of Rh antibodies.
Blood Types Compatibility
Red Blood Cells Compatibility Table

Donor
Recipient
Universal Recipient
Universal Donor
Hemolytic Disease of The Newborn (HDN)
 Also called, Erythroblastosis Fetalis a hemolytic anemia in the fetus or neonate, caused by trans-placental
transmission of maternal antibodies to fetal RBCs.
 Mother is Blood type Rh-, Father and fetus are Rh+.
 First pregnancy = Sensitization at delivery due to hemorrhage.
 Second pregnancy = Mother produce anti-Rh IgG antibodies that cross placenta to attack fetal RBCs leading to
hemolysis.
Practical Part 💉
Principle of Test
 The ABO and Rh blood grouping system is based on agglutination reaction.
 Agglutination is the reaction between antigens present on red blood cells and antibodies present in serum
resulting in visible clumping.
 Agglutination occurs if an antigen is mixed with its corresponding antibody, i.e. occurs when A antigen is mixed
with anti-A or when B antigen is mixed with anti-B.

Blood grouping solutions
Procedure
Results
 If the agglutination occurs in the RBCs to which both
anti-A and B is added, then the blood group is ‘AB’.
 If the agglutination occurs in the RBCs to which anti-
A is added, then the blood group is ‘A’.
 If agglutination occurs in the RBCs to which anti-B is
added, then the blood group is ‘B’.
 If there is no agglutination occurs in the RBCs, then
the blood group is ‘O’.
 If the agglutination occurs in the RBCs to which anti-D is added, then the blood type is positive (+) whereas if no agglutination
occurs in the RBCs to which anti-D is added, then the blood type is negative (-).

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Blood Biochemistry BCH 471[Practical]

Lab (3) ABO Blood Grouping & Rh Groups

1. To determine the blood group according to the ABO system.

2. To test for the availability of the Rh factor (D antigen).

Blood Group Substances Antigen ☓ Antibody

 ABO blood grouping consists of:

2. Two antibodies in the plasma (anti-A & anti-B)

Type AB A and B antigens No antibodies

 They are also useful in determining familial relationships in forensic medicine.

Red Blood Cells Compatibility Table

resulting in visible clumping.

with anti-A or when B antigen is mixed with anti-B.

 If the agglutination occurs in the RBCs to which both

anti-A and B is added, then the blood group is ‘AB’.

 If the agglutination occurs in the RBCs to which anti-

A is added, then the blood group is ‘A’.

 If agglutination occurs in the RBCs to which anti-B is

added, then the blood group is ‘B’.

 If there is no agglutination occurs in the RBCs, then

the blood group is ‘O’.

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