Download Molecular genotyping and frequencies of A , A , B, O and O

Int J Hematol (2008) 87:303–309
DOI 10.1007/s12185-008-0036-0
ORIGINAL ARTICLE
Molecular genotyping and frequencies of A1, A2, B, O1 and O2
alleles of the ABO blood group system in a Kuwaiti population
Mokhtar M. El-Zawahri Æ Yunus A. Luqmani
Received: 4 April 2007 / Revised: 26 November 2007 / Accepted: 14 December 2007 / Published online: 5 February 2008
Ó The Japanese Society of Hematology 2008
Abstract Expression of the highly polymorphic ABO gene
cluster is commonly investigated for blood transfusion and
analysis, but little information is available for Middle Eastern
populations. This study determined the major ABO allele
frequency in a Kuwaiti Arab cohort using a multiplex PCR–
RFLP technique; 355 unrelated blood donors of phenotype
A1 (46), A2 (31), A1B (6), A2B (4), B (97) and O (171) were
genotyped. DNA fragments of 252 (251 for O1) and 843 (842
for A2) bp spanning the two major exons, 6 and 7, of the ABO
gene were amplified and digested with HpaII and KpnI.
Thirteen different genotypes could be identified when combining the A1, A2, B, O1 and O2 alleles from the digestion
patterns: 1 A1A1 (0.28%), 6 A1A2 (1.69%), 38 A1O1
(10.71%), 1 A1O2 (0.28%), 1 A2A2 (0.28%), 30 A2O1
(8.45%), 6 A1B (1.69%), 4 A2B (1.13%), 12 BB (3.38%), 79
BO1 (22.25%), 6 BO2 (1.69%), 167 O1O1 (47.04%) and 4
O1O2 (1.13%). Two of the combinations (A2O2, O2O2) were
not found. All genotypes determined were consistent with the
serotypes. The frequencies of the five alleles in the Kuwaiti
sample population were ABO*A1 = 0.0746, ABO*A2 =
0.0592, ABO*B = 0.1676, ABO*O1 = 0.6831 and
ABO*O2 = 0.0155. These results are discussed with reference to gene frequencies reported for other ethnic groups.
Keywords Molecular genotyping Allele frequencies Suballeles ABO PCR–RFLP Kuwaiti population
M. M. El-Zawahri
Department of Biological Sciences, Faculty of Science,
Kuwait University, Safat, Kuwait
Y. A. Luqmani (&)
Department of Pharmaceutical Chemistry, Faculty of Pharmacy,
Kuwait University, P.O. Box 24923, 13110 Safat, Kuwait
e-mail: [email protected]
1 Introduction
Human population genetic studies were initiated following
the discovery of the ABO blood groups by Landsteiner in
1900 [1]. His findings of red cell agglutination by serum
and recognition of blood groups laid the scientific basis for
safe practice of blood transfusion. Red cell serology then
underwent a renaissance and the first examples of widespread human polymorphisms were defined. Recently, a
different form of classification of human blood groups has
been possible. More than 250 human blood group antigens
have been classified into 29 blood group systems by the
International Society of Blood Transfusion (ISBT) Working Party on Terminology for Red Cell Surface Antigens
[2]. The key features of systems and their antigens were
described by Reid and Lomas-Francis in 1997 [3].
Although dozens of blood systems have been identified, the
ABO blood group system has proven to be one of the most
widely analyzed polymorphisms, versatile and long lasting
in transfusion and transplantation medicine, forensic
serology, paternity testing, anthropologic investigation, and
personality prediction because of the inherent stability of
the antigens involved and the ability to readily obtain
results from dried blood stains [4, 5].
The ABO system involves four antigens: A, B, AB and
A1 [6]. The sequence of oligosaccharides determines
whether the antigen is A, B, or A1. For A/B antigen synthesis to occur a precursor called H antigen must be
present. H antigen is the only antigen of Hh blood group
system [6]. In RBCs, the enzyme that synthesizes the
H antigen is encoded by the H locus on chromosome 19 at
q13.3 [7]. The H antigen is converted to A or B by a1 ? 3N-acetyl-galactosaminyl-transferase (A transferase) or
a1 ? 3-galactosyl-transferase (B transferase), respectively. Both the transferases use the same acceptor
123
304
substrate (H antigen) but different nucleotide-sugar donor
substrates. On the other hand, group O individuals lack
such transferase enzymes and, consequently, continue to
express the basic H structure constituting a solitary terminal fucose moiety. The major A, B and O alloantigens are
coded by the A, B and O major alleles, respectively, at the
ABO locus on the human chromosome 9 at 9q34.1–q34.2.
Until recently, methods for determining ABO types have
been restricted to the serological test designed to detect
antibody or antigen materials and have been widely
adopted for several decades [8]. In addition to some technical drawbacks, the scope of these various serological
assays for polymorphism detection is relatively limited [9]
and even improvements [10] in these traditional methods
can only detect ABO phenotypes.
In 1990, Yamamoto and his colleagues elucidated the
molecular genetic basis of the major alleles at the ABO
locus [11–13]. Since then there has been substantial progress in our understanding of ABO genes and the
functional significance of the common allelic polymorphism at this locus. The ABO locus spans over 18 kb and
consists of seven exons that result in a protein of 354 amino
acids [6]. The exons range in size from 26 to 688 bp. Most
of the coding sequence is included in exon 7 [12–14].
Many mutations affecting the variety and specificity of the
encoded transferases have been identified. The nucleotide
sequences of the A and B genes are highly homologous
(99%); the A and B transferase genes differ in seven base
substitutions (at nucleotide positions 297, 526, 657, 703,
796, 803 and 930), but only four of these (at 526, 703, 796
and 803) result in an amino acid change, while the substitutions 297, 657 and 930 are silent. The O and A
transferase genes are identical except for a single cytosine
deletion at position 261 [12, 13]. The loss of this single
nucleotide results in a frame shift, producing a premature
stop codon, leading to a truncated, enzymatically inactive
protein of 115 amino acids [15]. A second O allele (O2)
that lacks this deletion has been described [16, 17], which
has some of the B allele mutations and an additional
mutation (G802Agly ? arg), that presumably inactivates
the enzyme. The A2 allele has been characterized [18] by a
single base deletion in the coding sequence which results in
an additional domain at the C terminus.
Molecular cloning of the ABO gene and elucidation of
the molecular basis of its various alleles allow the direct
determination of the ABO genotypes without the need for
family study. Not only has ABO genotyping become possible, but it has also become feasible to genetically
engineer the activity and specificity of the enzymes.
Variety of assays for polymorphism detection at the DNA
level has essentially replaced the former serological assays
even for routine ABO typing for blood transfusion. Various
methods have been used in the light of the increasing
123
M. M. El-Zawahri, Y. A. Luqmani
allelic variations. Several PCR based techniques such as
RFLP which provided the first basis for studying genetic
polymorphism at the DNA level [19], allele-specific PCR
(AS-PCR) [20], denaturing gradient gel electrophoresis
[21], single-strand conformation polymorphism (SSCP)
[22], amplified product length polymorphism (APLP) [23],
and others, take advantage of altered restriction enzyme
recognition sites caused by substitutions to genotype the
ABO locus for the A, B and O alleles and sub-alleles.
Single nucleotide polymorphisms (SNPs), mainly within
exons 6 and 7, define seven classical alleles: A1, A2, B, O1,
O2 and O3 (A101, A102, A201, B101, O101, O201 and
O303, respectively) [24]. More than 100 ABO alleles have
been characterized to date and are defined by different
SNPs within both coding and non-coding sequences [24–
29]. The incidence of the various molecular genotypes of
the ABO major-alleles and sub-alleles in the world population has been studied in restricted ethnic groups.
Polymorphisms due to ethnic and/or phenotypic variations
have been reported, underlining the necessity to look at
more populations to determine the extent of human
variation.
Kuwait is a small Arab country located northwest of the
Arabian Gulf on the northeastern part of the Arabian
Peninsula and shares borders with Saudi Arabia and Iraq. It
is an oil-producing country that is highly urbanized. The
surface area spans 17,818 km2 with a population of
2.895 million (2006 census). The population comprises of
approximately 34.54% Kuwaitis and 65.46% expatriate
workers of various nationalities, the majority of whom are
Arabs and Asians. The Kuwaiti population is a relatively
young population that has roots originating from its
neighboring countries. In this paper we report the results of
our study looking at allele frequencies for five of the
known ABO polymorphisms in the Kuwaiti population (A1,
A2, B, O1 and O2).
2 Materials and methods
2.1 Blood samples
Peripheral venous blood samples were collected into
standard hematological EDTA vials from 355 random
unrelated healthy Kuwaiti donors at the Kuwait Central
Blood Bank (consent was given). The expatriate workers
were excluded to obtain a pure Kuwaiti population.
2.2 Blood group serology
ABO phenotypes of the samples were determined by the
conventional erythrocyte and serum blood grouping
procedures.
Genotyping of ABO alleles in Kuwait
305
2.3 DNA preparation
3.2 Blood group genotyping
DNA was extracted from EDTA blood using Puregene
DNA Isolation Kit (Gentra, Minneapolis, USA) or Generation Capture Column Kit. Purified DNA samples were
quantified by spectrometry and diluted to approximately
100 ng/ll.
DNA preparations from 355 blood donors were amplified
in the multiplex PCR system. The fragments amplified
cover the two major exons and constitute 91% of the
translated gene. PCR using primer pair mo-57/mo-46 gives
a fragment of 252 bp (251 in O1) (exon 6) that is digested
by KpnI into 87 and 164 bp. PCR using primer pair mo101/
mo-71 gives a fragment of 843 bp (842 in A2) (exon 7) that
is degraded by HpaII. The 843 (842)-bp fragment also
includes 125 bp of untranslated DNA at the 30 end of the
gene (61 bp in A2 alleles) due to the single base deletion
resulting in an abolished stop codon and consequently a
larger reading frame [18]. After simultaneous digestion
with HpaII and KpnI, a conclusive interpretation of the
expected fragmentation pattern obtained could be made in
the 355 cases. Theoretically, 15 possible different genotypes could be identified from the possible combinations of
the five alleles from the digestion patterns (Table 1).
Examples of the electrophoretic patterns of the PCR
products after digestion of Kuwaiti DNA samples are
shown in Fig. 1. Thirteen different combinations of the five
alleles A1, A2, B, O1 and O2 were found in the Kuwaiti
population. Two of the combinations (A2O2 and O2O2)
were not found. The distribution of phenotypes, genotypes
and allele frequencies are show in Table 2. All genotyping
results were in agreement with the serologically determined phenotypes. The observed genotypic numbers did
not differ significantly from those expected under the
assumption of Hardy–Weinberg equilibrium (v2 = 10.015,
2.4 ABO genotyping
Using essentially the technique developed by Olsson and
Chester [28], a multiplex PCR with four primers amplifying exons 6 and 7 of the ABO genes was performed. (Gene
amplifier PCR system 9600, Perkin Elmer, was used.)
Fragments of 252 (251 for O1) and 843 (842 for A2) bp
were amplified in the same tube using the following ABO
primers (Oswel, UK):
mo-46: 50 -CGGAATTCACTCGCCACTGCCTGGGTC
TC-30 ;
mo-57: 50 -CGGGATCCATGTGGGTGGCACCCTGC
CA-30 ;
mo-101: 50 -CGGGATCCCCGTCCGCCTGCCTTGCA
G-30 ;
mo-71: 50 -GGGCCTAGGCTTCAGTTACTC-30 .
For each reaction 2.5 pmol of each of primers mo-46 and
mo-57 and 4 pmol of each of primers mo-101 and mo-71
were mixed with 5 ll (500 ng) genomic DNA in a total
volume of 40 ll containing (in final concentration) 0.2 mM
dNTPs, 2 mM MgCl2 and 1 unit Taq polymerase in the
buffer supplied. Amplification was performed with an
initial denaturation at 94°C for 2 min and then 10 cycles of
denaturation at 94°C for 10 s, annealing at 63°C for 60 s
and extension at 72°C for 1 min followed by 40 cycles of
denaturation at 94°C for 20 s, annealing at 61°C for 30 s
and extension at 72°C for 1 min.
Multiplex PCR products (5 ll) were digested using
0.5 ll (5 U) of each of restriction endonucleases HpaII and
KpnI (Invitrogen, Cambridge, UK) in 23 ll containing
0.36 ll BSA (10 lg) and 2.3 ll of 109 buffer BRL React
4. Cleavage products were separated electrophoretically
using 4% agarose gel containing ethidium bromide (Sigma,
St Louis, MO, USA).
Table 1 Digestion patterns for the genotypes identifiable from
combinations of the five sub-alleles (A1, A2, B, O1 and O2)
Genotypes Digestion products (bp)
87 96 119 137 150 164 204 223 252 309
A1A1
*
*
A1A2
*
*
A1O1
*
A1O2
A2A2
A2O1
*
*
*
*
*
*
*
*
*
A2O2
*
*
*
A1B
*
*
*
2
*
*
*
*
*
*
*
*
*
*
AB
3 Results
BB
BO1
3.1 Phenotypes of the blood samples
BO2
The phenotypes of the 355 blood samples used in this study
were A1 (46), A2 (31), A1B (6), A2B (4), B (97) and O
(171).
*
*
*
*
O1O1
*
*
O1O2
*
*
O2O2
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
Thirteen of the fifteen possible types were identified in our samples
123
306
M. M. El-Zawahri, Y. A. Luqmani
df 15, P [ 0.90). The frequencies of the five alleles in the
Kuwaiti sample population are ABO*A1 = 0.0746,
ABO*A2 = 0.0592,
ABO*B = 0.1676,
ABO*O1 =
2
0.6831 and ABO*O = 0.0155. The overall allele frequencies of the three major alleles A, B and O are 0.1338,
0.1676 and 0.6986, respectively. When compared by chisquare test, these allele frequencies were comparable to
those calculated from 18,555 Kuwaiti donors taken from
Kuwait Central Blood Bank database.
4 Discussion
310
281/271
234
194
BO2
O1O2
A2A2
A1O2
A1A1
A1A2
BB
A1B
A1O1
BO1
O1O1
A2O1
A2 B
118
M
Fig. 1 Gel electrophoretograms of the 13 genotypes which could be
detected amongst the 335 Kuwaiti DNA blood samples tested using
multiplex PCR–RFLP as described in Sect. 2. Lane M shows
molecular size marker, U 9 174RFDNA/HaeIII ladder; this strip
belongs to the same gel as the one shown for O1O1 and is included as
an indication only for the fragment sizes
Table 2 Phenotypes and frequencies of detected genotypes of the
ABO locus using a multiplex PCR–RFLP method in 355 blood
samples from Kuwaiti donors
ABO
ABO
Observed
phenotype genotype
Number
detected
A1
1
(0.28)
1.98
(0.56)
6
(1.69)
3.14
(0.88)
1
AO
38
(10.71)
36.18
(10.19)
A1O2
1
(0.28)
0.82
(0.23)
A2A2
1
(0.28)
1.24
(0.35)
A2O1
30
(8.45)
28.71
(8.09)
A2O2
0
0.65
(0.18)
A1B
A2B
2
B
(0)
Population
Allele frequencies
p
A
q
B
Reference
O
r
(1.69)
8.88
(2.5)
AB
4
(1.13)
7.04
(1.98)
Kuwaiti
0.1338
0.1676
0.6986
BB
12
(3.38)
9.97
(2.81)
BO1
Saudi Arabian
0.1663*
0.1197*
0.7140*
[30]
79
(22.25)
81.29
(22.9)
Bahraini
0.1410
0.1570
0.7040
[31]
6
(1.69)
1.84
(0.52)
Iraqi
0.2120*
0.1770
0.6611*
[32]
(46.66)
1
Present study
1
167
O1O2
4
(1.13)
7.52
(2.12)
O2O2
15
Iranian
Lebanese
0.2223**
0.3326**
0.1695
0.0996**
0.6082*
0.5680**
[33]
[34]
0
355
(0)
(100)
0.09
355
(0.02)
(100)
Palestinian
0.2790**
0.1250*
0.5930*
[35]
Jordanian
0.2700**
0.1300*
0.6000*
[36]
Egyptian
0.1880*
0.1490*
0.6630
[37]
Sudanese
0.1920**
0.1400*
0.6680*
[37]
OO
(47.04)
165.65
Allele frequencies: ABO*A1 = 0.0746; ABO*A2 = 0.0592;
ABO*O2 = 0.0155
ABO*B = 0.1676;
ABO*O1 = 0.6831;
(v2 = 10.015, df 15, P [ 0.90). Two of the combinations (A2O2,
O2O2) were not found
123
Table 3 Allele frequencies of the ABO gene in Kuwaitis in comparison with neighboring populations
6
BO2
Total
Number Frequency
(%)
A1A2
A1B
O
Frequency
(%)
A1A1
1
A2
Expected
This is the first report on the frequency of A2 and O2 alleles
in Arabian countries. The proposed study was designed so
as to provide insight on the molecular genetic profile of A1,
A2, B, O1 and O2 alleles of the ABO locus in 355 Kuwaiti
healthy donors using a multiplex PCR–RFLP technique
[28]. This method would miss other minor A or B genotypes. Genotyping was performed initially using PCR and
restriction enzyme analysis based on the mutation at
nucleotide 261 (O vs. A and B) and nucleotide 703 (A vs. B)
[12]. This was subsequently expanded to include detection
of the C526G mutation as suggested by Grunnet et al. [17]
to detect the presence of the O2 allele. The assignment of
the A2 allele was based on the C467T mutation [18] and
nucleotide 1060 deletion [28]. All allelic combinations
were observed in this group except A2O2 and O2O2. This
observation is not unusual given the frequency of these
alleles and the size of the study sample. The frequencies of
the five alleles within the study Kuwaiti sample population
were ABO*A1 = 0.0746, ABO*A2 = 0.0592, ABO*B =
0.1676, ABO*O1 = 0.6831 and ABO*O2 = 0.0155. The
highest frequency is O1 followed by B, A1, A2 and O2,
respectively. Unfortunately, there are no complete reports
about the frequencies of these five alleles in the neighboring populations from which the Kuwaiti population is
presumed to have originated. However, the frequencies of
the three major alleles (pA, qB and rO) calculated from the
results of the present study are compared with published
* P \ 0.05; **P \ 0.01
Genotyping of ABO alleles in Kuwait
307
data [30–37] on populations from neighboring countries
(Table 3). The studied Kuwaiti population was evidently
distinguished from the neighboring populations from which
it was originated. There is a statistically significant difference in allele frequency distribution between the
Kuwaiti population and all of the neighboring countries
except Bahrain. The frequencies of the three alleles in both
Kuwaiti and Bahraini populations are in the order of
rO [ qB [ pA, whilst the order is rO [ pA [ qB for the
remaining neighboring countries (Table 3). From a purely
scientific point of view, chemical analysis of the group O
antigen reveals that from a structural perspective it is the
simplest blood group and it serves as the backbone for the
synthesis of increasingly complex A, B and AB. These
latter blood groups evolved by adding other sugar onto the
basic O sugar. Thus if the mutations that produced the A
and B antigens are ancient, the gene for blood group O is
infinitely older. Among the Nomads of the Arabian Peninsula, and the Berbers of the Atlas Mountains, two old
populations, the frequency of the blood group O gene is
high. Africans, on average, have more O genes and less A
genes than do Europeans [4]. So it can be seen that the gene
carried by people who are blood group O is ancient by
evolutionary standards. Another interpretation regarding
the evolution of ABO antigen is that AB antigen was
already present in primates [38] and human ABO polymorphisms are probably selected by susceptibility to
certain infections.
The similarity between Kuwaiti and Bahraini populations may be explained on the basis of their similar
development. To obtain a clear picture of the Kuwaiti
population, one must consider its origin and background.
The Kuwaiti population is a relatively young population
that has roots originating from its neighboring countries.
Kuwait was first inhabited in the mid 1800s by settlers
coming from the Arabian Peninsula. During the 1900s,
more and more settlers came to Kuwait expanding its
population at a rapid rate. The new settlers originated not
only from the Arabian Peninsula but also from Iraq and
Iran further diversifying the genetic pool. Accordingly, the
Table 4 Frequencies of A1, A2,
B, O1 and O2 alleles of the ABO
gene in the Kuwaiti population
in comparison with some
worldwide populations
Population
Kuwaiti population is composed of multiplex Kuwaiti
families from three major Asian ancestral populations
(from Saudi Arabia, Iraq and Iran) and other minor Asian–
African ancestral populations (from Lebanon, Syria, Palestine, Jordan, Egypt and India). The nucleus population of
the Kuwaiti State, then, originated from small subpopulations forming a new heterogeneous population. This new
population has increased rapidly over the last five decades
from 0.299 million to nearly 1 million Kuwaiti nationals.
Due to the homogeneity and close social structure, each
individual appears to be exposed to a relatively common
environment. Therefore, the present study shows that there
are ethnic variations distinguishing this population from
other populations in neighboring countries with similar
ethnic background as well as from other populations.
Table 4 shows a comparison between the allelic frequencies of A1, A2, B, O1 and O2 in the Kuwaiti population
and some worldwide populations [28, 39–44]. The frequency of A2 allele in the Kuwaiti population is 5.92%,
which is similar to the European percentage (6.2%) and
significantly more than the Caucasian percentage
(1.888%). A2 alleles are mainly found in Europe and the
Near East and Africa [45]. This overall allele frequency
usually does not exceed 10% in Caucasian populations and
is very low (\1%) in Oriental populations [24, 40]. O1 and
O2 alleles were found in 97.77 and 2.23%, respectively, in
individuals with phenotype O; with frequency of 1.55%
and therefore O2 is a rare allele in the Kuwaiti population.
This frequency is similar to the Caucasian [41] and English
[44] populations. Reports described that the O2 allele
occurred at a frequency of 3.7% in Danish [17] and 3.33%
in Swedish [28] populations. In Chinese [39] and Japanese
[40] population O2 was not detected.
Molecular genotyping methods have several advantages.
This genomic DNA typing method provides a reliable
result. Since ABO blood group recording is very prevalent,
this approach obviously increases the value of this blood
group system in forensic suspect screening, paternity testing and anthropologic investigation [4, 5]. This method can
also help to elucidate the basis of unusual ABO blood
Reference number
ABO alleles
A1
A2
B
O1
O2
Kuwaitis (n = 335)
Present study
0.0746
0.0592
0.1676
0.6831
0.0155
Chinese (n = 315)
[39]
0.1920
0
0.1680
0.6300
0
Japanese (n = 340)
[40]
0.2660
0
0.1900
0.5400
0
Caucasian (n = 170)
[41]
0.1880
0.0170
0.1080
0.6710
0.0170
European (n = 300)
[42]
0.2150
0.0620
0.1120
0.5830
0.0280
German (n = 169)
[43]
0.2130
0.0770
0.0473
0.6420
0.0207
English (n = 86)
[44]
0.1980
0.0750
0.1050
0.6050
0.0170
Swedish (n = 150)
[28]
0.1900
0.0733
0.1033
0.5967
0.0333
123
308
groups such as cis-AB and Bombay phenotypes or variant
O alleles. In cis-AB phenotype, A and B genes are inherited simultaneously on one chromosome resulting in an
offspring inheriting three ABO genes instead of two [46].
In individuals of the Bombay phenotype, the red cells and
secretions express no A, B, or H antigens, although A and/
or B alleles are present. This uncommon Bombay phenotype is a result of homozygosity for the h allele (h/h) [47].
In addition, this approach can determine the ABO genotype
of blood group changes in malignancy [48]. Finally, the
applications of this method in organ transplantations have
lead to safer and more reliable ways of improving the
compatibility of donors with ethnic disparity [49].
Acknowledgments This work was supported by grant SZ 01/00
from Research Administration, Kuwait University, Kuwait. The
authors are indebted to Dr Abdul-Aziz Al-Bashir (Director of the
Central Blood Bank of Kuwait) for facilitating collection of the blood
donor samples and Ms Gilu Abraham for her technical assistance.
References
1. Landsteiner K. Ueber agglutinationsercheinungen normalen
menschichen blutes. Wein Klin Wochenschr. 1901;14:1132–4.
2. Reid M, McManus K, Zelinski T. Chromosome location of genes
encoding human blood groups. Transfus Med Rev. 1998;3:151–
61.
3. Reid M, Lomas-Francis C. The blood group antigen facts. NY:
Academic Press; 1997.
4. Mourant A, Kopec A, Domaniewska K. The distribution of the
human blood groups and other polymorphisms. Oxford: Oxford
University Press; 1976.
5. Lee H. Identification and grouping of blood stains. In: Saferstain
R, editor. Forensic science handbook. Englewood Cliffs: Prentice
Hall; 1982.
6. Reid ME, Lomas-Francis C. The blood group antigen facts book.
2nd ed. New York: Elsevier Academic Press; 2004.
7. Kelly RJ, Ernst LK, Larsen RD, Bryant JG, Robinson JS, Lowe
JB. Molecular basis for H blood group deficiency in Bombay
(Oh) and para-Bombay individuals. Proc Natl Acad Sci USA.
1994;91:5843–7.
8. Boulton S, Thorpe J. Enzyme-linked immunosorbent assay for A
and B water soluble blood group substances. J Forensic Sci.
1986;31:27–35.
9. Race R, Sanger R. Blood groups in man. London: Black well
Scientific Publications; 1975.
10. Zhou B, Gou J, Wang C, Chen J. The rapid determination of
ABO group from body fluid (or stains) by dot enzyme-linked
immunosorbent assay (Dot-ELISA) using enzyme-labeled
monoclonal antibodies. J Forensic Sci. 1990;35:1125–32.
11. Yamamoto F, Marken J, Tsuji T, White T, Clausen H, Hakomori
S. Cloning, characterization of DNA complementary to human
UDP-GalNAC: Fuca1–2Gal a1- 3GalNAc transferase (histoblood
group A transferase) on RNA. J Biol Chem. 1990;265:1146–51.
12. Yamamoto F, Clausen H, White T, Marken J, Hakomori S.
Molecular genetic basis of the histo-blood group ABO system.
Nature. 1990;345:229–33.
13. Yamamoto F, Hakomori S. Sugar-nucleotide donor specificity of
histo-blood group A and B transferases is based on amino acid
substitutions. J Biol Chem. 1990;265:19257–62.
123
M. M. El-Zawahri, Y. A. Luqmani
14. Yamamoto F. Molecular genetics of the ABO histoblood group
system. Vox Sang. 1995;69:1–7.
15. Lutz P, Dzik W. Molecular biology of red cell blood group genes.
Transfusion. 1992;32:467–83.
16. Yamamoto F, McNeil P, Yamamoto M, Hakomori S, Bromilow,
IDuguid K. Molecular genetic analysis of the ABO blood group
system: 4. Another type of O allele. Vox Sang. 1993;64:175–8.
17. Grunnet N, Steffensen R, Bennet E, Clausen H. Evaluation of
histo-blood group ABO genotyping in a Danish population: frequency of a novel O allele defined as O2. Vox Sang.
1994;67:210–5.
18. Yamamoto F, McNeill P, Hakomori S. Human histo-blood group
A2 transferase coded by A2 allele, one of the A subtypes, is
characterized by a single base deletion in the coding sequence,
which results in an additional domain at the carboxyl terminal.
Biochem Biophys Res Commun. 1992;187:366–74.
19. Lee J, Chang J. ABO genotyping by polymerase chain reaction.
J Forensic Sci. 1992;37:1269–75.
20. Ugozzoli L, Wallace R. Application of an allele-specific polymerase chain reaction to the direct determination of ABO blood
group genotypes. Genomics. 1992;12:670–4.
21. Jonhson P, Hopkinson D. Detection of ABO blood group polymorphism by denaturing gradient gel electrophoresis. Hum Mol
Genet. 1992;1:341–4.
22. Akane A, Yoshimura S, Yoshida M, Okii Y, Watabiki T,
Matsubara K, et al. ABO genotyping following a single PCR
amplification. J Forensic Sci. 1996;41:272–4.
23. Watanabe G, Umetsu K, Yuasa I, Suzuki T. Amplified product
length polymorphism (APLP): a novel strategy for genotyping the
ABO blood group. Hum Genet. 1997;99:34–7.
24. Yip S. Sequence variation at the human ABO locus. Ann Hum
Genet. 2002;66:1–27.
25. Olsson M, Chester M. Polymorphism and recombination events
at the ABO blood locus: a major challenge for genomic ABO
blood grouping strategies. Transfus Med. 2001;11:295–313.
26. Hosseini-Maaf B, Hellberg A, Rodrigues M, Chester M, Olsson
L. ABO exon and intron analysis in individuals with the AweakB
phenotype reveals a novel OIv-A2 hybrid allele that causes four
missense mutations in the A transferase. BMC Genet. 2003;4:17–
27.
27. Yamamoto F. Blood group antigen gene mutation database—
ABO blood group system, common alleles of ABO locus.
http://www.bioc.aecom.yu.edu/bgmut/abo_common.htm. Accessed 7 Nov 2004.
28. Olsson M, Chester M. A rapid and simple ABO genotype
screening method using a novel B/O2 versus A/O2 discriminating
nucleotide substitution at the ABO locus. Vox Sang.
1995;69:242–7.
29. Olsson M, Thuresson B, Chester M. An Ael allele-specific
nucleotide insertion at the blood group ABO locus and its
detection using a sequence-specific polymerase chain reaction.
Biochem Biophys Res Commun. 1995;216:642–7.
30. Bashwari L, Al-Mulhim A, Ahmad M, Ahmed M. Frequency of
ABO blood groups in the eastern region of Saudi Arabia. Saudi
Med J. 2001;22:1008–12.
31. Al-Arrayed S, Shome D, Hafadh N, Amin S, Al-Mukhareq H, AlMulla, et al. ABO blood group and Rhd phenotypes in Bahrain:
Results of screening school children and blood donors. Bahrain
Med Bull. 2001;23:112–5.
32. Tills D, Kopec A, Tills R. The distribution of the human blood
groups and other polymorphisms. Supplement 1. Oxford/New
York/Toronto: Oxford University Press; 1983.
33. Walter H, Farhud D, Danker-hopfe H, Amirshahi P. Investigations on the ethnic variability of the ABO blood group
polymorphism in Iran. Z Morphol Anthropol. 1991;78:289–306.
Genotyping of ABO alleles in Kuwait
34. Kfoury E, Mahfouz R, Tabarani R, Ayoub T, Zaatari G. Lebanese
population: prevalence of the erythrocyte phenotypes. Leban Med
J. 2001;49:140–2.
35. Dudin A, Rambaud-Cousson A, Badawi S, Dana N, Malji A,
Hannoun A. ABO and Rh(D) blood group distribution and their
implication for feto-maternal incompatibility among the Palestinian population. Ann Trop Paediatr. 1993;13:249–52.
36. Irshaid N, Ramadan S, Wester E, Olausson P, Hellberg A, Merza
JY, et al. Phenotype prediction by DNA-based typing of clinically
significant blood group systems in Jordanian blood donors. Vox
Sang. 2002;83:55–62.
37. Khalil I, Phrykian S, Farri A. Blood group distribution in Sudan.
Gene Geogr. 1989;3:7–10.
38. Martinko JM, Vincek V, Klein D, Klein J. Evidence for transspecies evolution. Immunogenetics. 1993;37:274–8.
39. Yip S. Sequence variation at the human ABO locus. Ann Hum
Genet. 2000;66:1–27.
40. Fukomori Y, Ohnoki S, Shibata H, Nishimukai H. Suballeles of
the ABO blood group system in a Japanese population. Hum
Hered. 1996;46:85–91.
41. Pearson S, Hessner M. A1,2, B, O1,2 genotyping by multiplexed
allele-specific PCR. Br J Haematol. 1998;100:229–34.
42. Gassner C, Schmadra A, Nussbaumer W, Schonitzer D. ABO
glycosyltransferase genotyping by polymerase chain reaction
using sequence specific primers. Blood. 1996;88:1852–6.
309
43. Nishimukai H, Okiura T, Shinomiya T, Fukumori Y, Ohnoki S,
Shibata H, et al. Genotyping of the ABO blood group system:
analysis of nucleotide position 802 by PCR–RFLP and the distribution of ABO genotypes in a German population. Int J Legal
Med. 1996;109:90–3.
44. Procter J, Crawford J, Bunce M, Welsh K. A rapid molecular
method (polymerase chain reaction with sequence-specific
primers) to genotype for ABO blood group and secretor status
and its potential for organ transplants. Tissue Antigens.
1997;50:475–83.
45. Daniels G. Human blood groups. Cambridge: Blackwell Science;
1995.
46. Wallace EM, Gibbs LF. Blood group systems: ABH and Lewis.
Virginia: American Association of Blood Banks; 1986.
47. Mateos P, Caillean A, Henry S, Costache M, Elmgren A,
Svensson L, et al. Point mutations and deletion responsible for
the Bombay H null and the reunion H weak blood groups. Vox
Sang. 1998;75:37–46.
48. O’Keefe SD, Dobrovic D. A rapid and reliable PCR method for
genotyping the ABO blood group. Hum Mutat. 1996;2:67–70.
49. Haji NI, Nelson RE, Jones MN, Mark NA, Johnson RJ, de Silva M,
et al. Will transplantation of kidney from donors with blood group
A2 into recipients with blood group B help British Indo-Asian
patients with renal failure? Transplantation. 2004;77:630–3.
123