BASICS OF ABO

P.Renuka
Objectives
-History
- Inheritance of ABO blood groups
- ABO sub groups
- Technique of grouping
- ABH antigens and antibodies in disease
- Recent advances
Only blood group system in which individuals have antibodies in their
serum to antigens that are absent from their RBCs
History
• A,B,O discovered by Lansteiner in 1901
• AB discovered by Adriano Sturli and Alfred von Decastello – who
were working under Landsteiner – discovered type AB a year
later in 1902
Inheritance
• Bernstein in 1924- an individual inherits one ABO gene from each
parent .Follows Mendelian genetics.
• Co dominant in expression.
• One locus on chromosome 9 is occupied by A,B or O gene.
• O gene is considered as AMORPH
• Designations A and B refer to phenotypes
• AA,BO and OO refer to genotype.
ABO genotypes and phenotypes

Prevalence of blood groups among Indians

makro
Formation of A,B and H red cell antigens
• Interaction of genes at three separate loci- ABO,Hh,Se
Inheritance of H gene results in H antigen.
H and Se genes are not part of the ABO system,they influence the A
and B antigen expression
H and Se genes – chromosome 19

• Produce specific glycosyl transferases that add sugars to basic

precursor substance.
• Paragloboside or glycan – precursor
• ABH antigens develop early in fetal life,do not increase in strength
during gestational period.
• New born – from 25% to 50% of number of antigenic sites.
• Fully developed by 2 to 4 years
blood group O
Inherit atleast one H gene [genotype HH or Hh] and two O genes.
H gene elicits production of alpha 2 L fucosyl transferase which
transfers sugar L fucose to an oligosaccharide on the terminal galactose
of type 2 chains on RBC membrane.
Immunodominant sugar
• Tab 6-9
• Fig 6-6
• B enzyme seems to compete more efficiently than A for the H
substance.
Molecular genetic of ABO
ABO gene consists of seven exons ,last two exons code for ABO glycosyl
transferase.
Amino acid substituitions resulting from mutation within these two
exons –less efficient transfer of immuno dominant sugar to H
substance.Results in weak serologic reactions observed in ABO sub
groups.
 A ,B and H soluble antigens
• ABH antigens are integral parts of membranes of RBCs,endothelial
cells,platelets,lymphocytes and epithelial cells.
• Found in all body secretions.Their presence is dependent on ABO
genes inherited through genes called Sese {secretor genes}
• Sese gene codes for the production of alpha 2 L –fucosyl
transferase,that modifies the type I precursor substance to form H
substance.
• People who inherit the sese genotype are termed non secretors.
ABO sub groups
• 1911 von Dungern described two different antigens
• Group A RBCs react with both anti –A and anti – A1 are classified as
A1,those react with anti A and not anti – A1 are classified as A2.
• Four different forms of H antigens – H1,H2,H3 and H4.
• Weak subgroups –A3,AX,Aend,Am ,Ay and A el.
Investigation of weak A subgroups
Weak B sub groups
• Very rare.Less frequent than A
• Include B 3,Bx,Bm and Bel.
• Table 6-16
The Bombay Phenotype(0h)
• First reported by BHENDE in 1952 in Bombay ,India.
• Autosomal recessive trait.Mutation in FUTI gene ,incapable of coding for
alpha 1,2 fucosyl transferase.
• Inheritance of double dose of h gene producing hh.As a result,ABO genes
not expressed,ABH antigens not formed,no H antigen made.
• More than 130 Bombay phenotypes reported in various parts of the
world
• Anti A ,anti –B,anti A,B and anti H present in the serum
• Can only be transfused with blood from another Bombay (Oh)
• PARA BOMBAY group – weak forms of A and B antigens(detected by
adsorption and elution test)
Technique of blood grouping
- phenotyping and genotyping
Phenotyping
• Slide technique
• Tube technique
• Microtitre plate technique
• Column agglutination technique

Genotyping
Micro titre plate

• Micro titre plate foto

• Gel card foto
Genotyping
Sample – white blood cells from peripheral blood
PCR -
• PCR RFLP - amplicon followed by digestion with restriction enzyme ,
Cleave unique sequence of nucleotide followed by gel electrophoresis.
Fragment pattern compared with known controls.
REAL TIME PCR – amplified DNA is detected during PCR.Non specific
fluorescent dyes added interact with DNA.
Micro array technology –uses colour coded beads coated with allele
specific oligo nucleotides.PCR product annealed to beads,hybridized
product match or mismatch.Match – fluorescent signal –decoded by
software.Mismatch – no signal.
Diseases and blood groups
Diseases alter red cell antigens
Leukemia, hemolytic disease ,hodgkins disease depress antigen
strength-mixed field agglutination
Iso agglutinins anti – A, anti –B or anti A,B weak or absent in
hypogammaglobulinemia,CLL, Non hodgskins lymphoma.
Acquired B phenomenon in intestinal obstruction,carcinoma of colon or
rectum
Lack of antigens in carcinoma of stomach or pancreas.
Transfusion journal Nov 2018
Use of low titre group O whole blood for resuscitation of civilian trauma
patients in 2018
ABO group unknown
Low titre O group whole blood must be utilized.The most common
definitions of low titre anti A and B was less than 200 and 256
Ig M titre of <250.IgG titre <500
Blood group p null
A team of doctors led by Dr.Sharma shastry from the blood bank of
Kasturba medical college ,Manipal identified a rare blood group called pp
or P null phenotype in July 2018
Crossmatching – could not find an unit.So referred to international
blood group reference laboratory ,Bristol,U.K for serological study.

Incidentally Department of immunohaematology under ICMR Mumbai is

our national level reference laboratory for blood group genomics in India.