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ISSN 2348 – 0319
International Journal of Innovative and Applied Research (2014), Volume 2, Issue (8): 57- 60
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INTERNATIONAL JOURNAL
OF INNOVATIVE AND
APPLIED RESEARCH
RESEARCH ARTICLE
ABO and Subgroups phenotype in Iraqi Paternity Cases
Jawad. k. Mashaali
M.Sc. Genetic Engineering and Biotechnology
………………………………………………………………………………………………………
Abstract:
Background.
Traditional ABO blood group serology is based on the immunoreactivity of antisera with the carbohydrate A , B
and H antigens on Red Blood Cells ( RBC) . Progress in the molecular biology of the ABO system has recognized
the molecular basis of the red blood cell antigens and has provided a genetic model for ABO polymorphism at the
molecular level. The ABH antigens are not primary gene product but they are the enzymatic reaction product of
enzymes called glycosyltransferases.
Aim of study.
The study attempted to document the frequency of the ABO and rhesus blood group phenotype including the C,
E, e, c antigens with the possible genotypes in our cases.
Method .
Samples of 100 randomly healthy Iraqi voluntary blood donors were collected from the Iraqi National Blood
Bank, Baghdad. the sample were used for estimation of phenotypic distribution of ABO , Rh and subgroup E, e,C, c
. ABO blood group serology was performed at medico-legal institute ,Paternity and Kinship Division using
commercially available monoclonal Anti-A, Anti-B , reagents according to the national blood bank standard
operating procedures (1) .Method principle is based on antigen antibody reaction ( agglutination ) some tests were
confirmed under microscope examination to see weak reaction .
Key Words: Antigens ,ABO system , Rh system , E , e , C , c , subgroup.
………………………………………………………………………………………………………
Introduction
Experiments with blood transfusions have been carried out for hundreds of years. Many patients have died and it
was not until 1901, when the Austrian Karl Landsteiner discovered human blood groups, that blood transfusions
became safer.He found that mixing blood from two individuals can lead to blood clumping. The clumped RBC( Red
Blood Cells ) can crack and cause toxic reactions, this can be fatal (2) .
Karl Landsteiner discovered that blood clumping was an immunological reaction which occurs when the
receiver of a blood transfusion has antibodies against the donor blood cells (3).The blood group antigen are of clinical
importance in blood transfusion , organ transplantation , autoimmune haemolytic anaemia , fetomaternal blood
group incompatibility, paternity identification , anthropologic investigation and forensic medicine . There are 29
systems for blood grouping classification. One of these is the ABO system (4).The majority of cell types investigated
in human have A , B , and O antigen originally found on the surfaces of red cells (5) , but later they were also found
on surface of various types of cells as well as in secretions. This include platelets , lung tissues , intestinal mucosa ,
mucous cells , epidermis , nervous receptors and vascular endothelium (6,7).
In 1910, scientists proved that the RBCs antigens were inherited, and that the A and B antigens were inherited co
dominantly over O . There was initially some confusion over how a person's blood type was determined, but the
puzzle was solved in 1924 by Bernstein’s” three allele model”. The ABO blood group antigens are encoded by one
genetic locus, the ABO locus, which has three alternative (allelic) forms A, B, and O.
A child receives one of the three alleles from each parent, giving rise to six possible genotypes and four possible
blood types (phenotypes).The ABO antigens are not the primary gene products, but instead the enzymatic reaction
product catalyzed by the enzymes called glycosyltransferases , and the final step of their synthesis is catalyzed by A
or B transferees which is encoded by the functional alleles ( A or B , respectively) at the ABO locus . Group A and
B glycosyltransferases are responsible for the conversion of H antigen to A and B antigen. The genes that encode the
transferees responsible for these reactions have been cloned and sequenced in 1990.
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International Journal of Innovative and Applied Research (2014), Volume 2, Issue (8): 57- 60
Genotyping enables the identification of both maternally and paternally derived alleles. Due to large number of
alleles that could give similar phenotype but differ in genomic structure, there are several reasons for determining
genotyping for blood group. These reasons include forensic medicine and paternity testing (8,9,10) and linkage
analysis (11) .Conventional serological method problems are false positive or negative and low antibody titer of
antisera.
Several new strategies have been employed to detect ABO genotype and other genetic markers by PCR and
DNA typing. Restriction fragment length polymorphism (RFLP) method has taken advantage of altered restriction
enzyme recognition sites caused by nucleotide substitution in the ABO locus of the A , B, or O alleles .This method
used to differentiate between B and O alleles by PCR-RFLP (12,13,14,15). In Iraq there are two specific studies for
ABO system one at phenotypic level which revealed that: blood group O is the most common (35.7%) followed by
B (28.3%), A (26.2%) and AB (9.8%).Rh D blood grouping was also evaluated and showed that; Rh D positive
blood were (92%), while Rh D negative blood were (8%), (16) .the second study at molecular level which revealed
the thirty samples show 14 ( A )blood group, 13 of them are Heterozygous AO and one is Homozygous AA . The
14 samples are blood group B, 13 of them are Heterozygous BO and one of them is Homozygous BB. One sample
for AB, O blood groups this type have only Homozygous phase and each gene are dominant in AB type so called
co-dominant. In O type the gene is recessive and expressed only in homozygous type only whereas the A and B
genes are dominant over O, (17).
In 1940, Landsteiner and Wiener found that sera from rabbits (and later guinea pigs) immunized with RBCs
from rhesus monkeys red cells agglutinated 85% of human RBC samples (18).Initially, it was thought that the animal
and human antibodies identified a common factor, Rh, on the surface of rhesus and human RBCs (19).The heteroantibody was renamed anti-LW after (Landsteiner and Wiener), and the human allo-antibody was renamed anti-D.
Material and methods
Conventional blood group ( ABO) and subgroup E,e,C,c were done for 100 donors , blood samples collected in
EDTA tubes. The BioTech kit, for ABO , Rh and E , C, e , c , subgroup were used . Slid method was done for this
purpose (20). the weak antigen may be need exam under microscope . The difference in amount of antigen expressed
on the red cell membrane between a homozygous and a heterozygous can often be detected serologically and is
termed the dosage effect. The result of agglutination classified to about four grades according to density of
agglutination. (21).
Discussion
The terminologies used to describe the Rh system are derived from four sets of investigators. Two of the
terminologies are based on the postulated genetic mechanisms of the Rh system. The third terminology described
only the presence or absence of a given antigen. The fourth is the result of the combined efforts of the International
Society of Blood Transfusion (ISBT) (22). In the early 1940, Fisher and Race postulated the antigens of the system
were produced by three closely linked sets of alleles. Each gene was responsible for producing a product (or antigen)
on the red cell surface. Fisher and Race named the antigens of the system D, d, C, c, E, and e. To date, no d antigen
has been found and it is considered an a morph (silent allele) or the absence of D antigen (23), (Figure .1).
D gene
Close
Production
C/c gene
Linkage
Antigens
D
C/c
Pathway
E/e gene
E/e
Figure ( 1): Fisher-Race concept of Rh (simplified).
Each gene produces one product (Denise, 2001)
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International Journal of Innovative and Applied Research (2014), Volume 2, Issue (8): 57- 60
The common Rh antigens D, C or c and E or e, were originally written in alphabetical order (CDE) but later,
when it was recognized that C and E antigens are inherited enbloc, the order was changed to DCE. Fisher postulated
that the C/c locus lies between D/d and E/e loci. This information is based on frequencies of the various gene
combinations (1).
Result
ABO and Rh system with E , e , C , c , are all somatic genes traits , therefore no effected to gender, birth place,
age, and race on those traits , never effected to general ratio of all blood group because no effected to environment
on expression of gene to this trait , this called pure genetic trait which controlled by gene only rather than other traits
which controlled by gene and environment like tall and skin colour .
The gene of ABO is independent to RH gene each one of them had specific sequence on DNA ( gene ) , therefore
each of them inherited separated for them, but there some neighborhood ( unblocked genes) between RH gene and E
, e . C, c genes therefore there were high inherited significant between them.
There are high significant between birth place and race because the person or donors are had the same address
and born place (Baghdad).
Glass method were used ,the results of the phenotypic distribution indicated that 100 of persons blood were
revealed that as showed in Table(1) blood group O is the most common (39%) followed by A(26%) , B ( 24% ) and
AB (11%).
Rh blood grouping was also evaluated and showed that; Rh positive blood were (80%) while Rh negative blood
were (20%).
The results revealed that high significant between RH and E, C followed by clear relation between (E subgroup and
e, c subgroup), and C subgroup and e, c subgroup.
There are no relationship between the following parameter, sex, age, Birth place, Race and Rh with ABO blood
group.
The results of ABO, Rh, E, e, C, c are shown in table (1).
Bl. group
And
Subgroup
Number
positive
negative
A
26
B
24
AB
11
O
39
RH
80
20
E
32
68
e
94
6
C
77
23
67
33
c
Total
100
Male
96
Female
4
References
1- Daniels, G.L ( 1995 ).Modern blood banking and transfusion practices 4th Edition, Oxford, Blackwell Science .
2- Eastlurd, T(1998).”The Histo-blood Group ABO System and Tissue Transplantation”. Transf; 38: 975-988.
3- Reid ,M.E and Lomas-Francis ,C (2004).The blood group antigen fact book 2 nd Edition ,New York; Elsevier
Academic Press.
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ISSN 2348 – 0319
International Journal of Innovative and Applied Research (2014), Volume 2, Issue (8): 57- 60
4- Daniels,G.L(2007) ”Red Cells Blood Grouping in the Molecular Era”Vox Sang;2:53-58.
5- Race, R . and Sanger, R( 1975). Blood Groups in Man. Blackwell scientific publication. Oxford .UK.
6- Orial, R(1992). ”ABO, H, Lewis and Secretion: Serology, Genetics and Tissue Distribution” . In: Blood cell
biochemistry, vol.VI: Molecular Basis of Major Human Blood Group Antigens, Plenum, New York, 37-73.
7- Brian , R.C.;Edwards ,M.J.;Hessner,J.P.;Klein and Aster,R.H(2000 ).” Blood group A and B antigens are strongly
expressed on platelets of some individuals”. Blood ;96:1574-1581.
8-Chester,M.A. and Olsson,M.L(2001).”The ABO Blood Ggroup Gene: A locus of considerable genetic diversity”.
Transf .Med.Rev;15:177-200.
9-Hashimoto,Y.; and Nakanishi,A(1993).”ABO Genotyping by Polymerase Chain Reaction and its Application to
Paternity Testing” .Nippon Hoigaku Zasshi;47:481-465 .
10-Sasaki,T.;Fukushima,M and Shiono,H(1994).”ABO genotyping of Fingerprints by The PCR-RFLP Method”.
Nippon Hoigaku Zasshi;48:428-432.
11-Yip,S.P.;Yow,M.C and Lewis,W.H (1995).”DNA Polymorphism at The ABO Locus in The Chinese Population
of Hong Kong” .Hum.Hered;45:266-271.
12-Yamamoto ,F.;Clausen , T .;White ,L.;Marken and S.Hakomori (1990b). Molecular Genetic Basis of The Histoblood Group ABO System”.Nature; 345: 229-233 .
13-Lee ,J.C and Chang,J.G( 1992 ).”ABO Genotyping by Polymerase Chain Reaction”. J .Forensic Sci ;37:12691275 .
14-O’Keefe ,D.S. and Dobrovic,A(1996).”A Rapid and Reliable PCR Method for Genotyping The ABO Blood
Group Alleles”.Hum.Mutat;8:358-361.
15-Stroncek , D.F.;R.Konz ,M.E.;Clay ,J.P.;Houchins and Ullough,J( 1995). ”Determination of ABO
Glycosyltransferase Genotypes by Use Polymerase Chain Reaction and Restriction Enzymes” .Transf;35:231 -240 .
16- Rahim, Alyaa.A (2005).”ABO and Rh blood groups with Rh phenotypes in Iraq”. Master thesis, University of
Baghdad .College of medicine. Hematology.
17- Mohammed, Ahmed .K(2008) . "Molecular detection of blood group ABO by PCR-RFLP method "Master
thesis , University of Baghdad . Genetic engineering and biotechnology institute for postgraduate studies .
18 - Mollison PL: The genetic basis of the Rh blood group system. Transfusion 1994; 34: 539-5191.
19 - Cartron JP. Agre P. Rh blood group antigens protein and gene structure. Seminars in Hematology 1993; 30:
193-208.
20 - Daniels, G.L ( 1995 ).Modern blood banking and transfusion practices 4 th Edition, Oxford, Blackwell Science .
21- Boorman,K.E and Dodd,B.E(1961).An Introduction to Blood Group Serology .Theory ,Techniques, Practical
Applications. 3rd Edition, Churchill Livingstone.UK.
22 - Issitt PD, Crcokston MC, Blood group terminology : Current conventions. Transfusion, 1984; 24:1.
23 - Lewis M, Anstee DJ, Bird; GWG. et al. Blood group terminology, from the ISBT Working party on
terminology for red cell surface antigens. Vox Sang, 1990; 58: 152.
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