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Red blood cell phenotyping of blood donors in Islamabad, Pakistan

Article  in  Global Journal of Transfusion Medicine · April 2018

DOI: 10.4103/GJTM.GJTM_54_17

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Original Article

Red Blood Cell Phenotyping of Blood Donors in Islamabad, Pakistan

Faiza Jabin1,2, Usman Waheed3, Saeed Ahmed4, Muhammad Arshad5, Abida Arshad2, Hasan A Zaheer6

1
Department of Pathology Background: There are 33 blood groups identified so far representing more

Abstract
and Blood Bank, Shaheed
Zulfiqar Ali Bhutto
than 300 antigens. The blood transfusion services in resource‑limited countries
Medical University (PIMS), including Pakistan only test for ABO and RhD antigens during cross‑matching.
2
Department of Zoology, However, the transfusion of ABO‑RhD compatible but unknown phenotype blood
PMAS Arid Agriculture may result in alloimmunization, especially in multi‑transfused patients. Aims: The
University, Rawalpindi, aim of this study is to investigate the prevalence of clinically significant minor
3
Safe Blood Transfusion blood group antigens in population of Islamabad, Pakistan. Settings and Design:
Programme, Ministry of
National Health Services,
A  prospective study was conducted at the Department of Blood Transfusion
Government of Pakistan, Services, Islamabad. Subjects and Methods: Blood samples from 625 randomly
4 
Department of Blood selected blood donors were collected for extended antigen typing. Blood grouping
Bank, Husaini Haematology was performed using commercially available antisera. The statistical analysis
and Oncology Trust, was done using SPSS version 20.0. Gene frequencies were calculated using the
Karachi, 5Department Hardy–Weinberg equation. Results: Of the 625 blood donors, 575 were RhD
of Bioinformatics and
Biotechnology, International
positive (92%) and 50 (8%) were RhD negative. The frequencies of A, B, AB,
Islamic University, and O antigens were (24.64%), (34.72%), (9.28%), and (31.36%), while antigen
6
Department of Pathology frequency of K was 4%, Duffy (Fya) 58.24%, Duffy (Fyb) 39.84%, Kidd (Jka)
and Blood Bank, Shaheed 65.28%, Kidd (Jkb) 42.24%, M 86.88%, N 46.24%, S s49.44%, and s 30.4%.
Zulfiqar Ali Bhutto Medical Calculated gene frequency for Kell K was (0.04), Duffy Fya (0.481), Fyb (0.519),
University, Islamabad, Kidd JKa (0.538), JKb (0.462), M (0.699), N (0.301), S (0.445), and s (0.556).
Pakistan
Conclusions: The current study depicts the prevalence of the blood group antigens
among Pakistani blood donors to be statistically different from those in Caucasians,
Chinese, and  African populations.
Received: October, 2017.
Accepted: January, 2018. Keywords: ABO, alloimmunization, Islamabad, red cell phenotypes

Introduction immunogenic and active at body temperature (37°C)[2]

H uman blood group systems are highly
polymorphic. Antigenic substances (proteins),
resided on the surface of erythrocytes, make the base
of different blood group systems. These antigens
are the inherited characters.[1] Over 300 blood group
antigens present on the surface of red blood cells[2]
and 33 blood groups have been recognized so far by
ISBT.[3] Transfusion therapy becomes complicated
due to alloantibodies and cause immediate or delayed
hemolytic transfusion reactions, with an incidence
of around 1/7000 RBC units transfused.[4] Apart
from major blood group systems (ABO and Rhesus),
minor blood groups including Kidd, Duffy, and Kell
blood group systems play substantial role in clinical
transfusion practices because their antigens are more
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In routine blood banking, blood transfusion services
are confined only to ABO and RhD antigen typing,
particularly in resource‑limited countries,[5] including
Pakistan. Normally, it is considered secure
and time‑saving, but for the multi‑transfused
patients (thalassemic and anemic), the extended
phenotyping for the determination of minor antigens is
also necessary. The frequencies of clinically significant
blood groups antigens (Duffy, Kell, Kidd, and MNS)
should be determined while dealing with patients who
have multiple alloantibodies.[6] Complications related
to blood transfusion can be avoided by having proper
Address for correspondence:
Dr. Usman Waheed, E-mail: usman.waheed07@gmail.com
This is an open access journal, and articles are distributed under the terms of the Creative
Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to
remix, tweak, and build upon the work non-commercially, as long as appropriate credit is
given and the new creations are licensed under the identical terms.
For reprints contact: reprints@medknow.com

How to cite this article: Jabin F, Waheed U, Ahmed S, Arshad M,

DOI: 10.4103/GJTM.GJTM_54_17 Arshad A, Zaheer HA. Red blood cell phenotyping of blood donors in
Islamabad, Pakistan. Glob J Transfus Med 2018;3:26-9.

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Jabin, et al.: Red Blood Cells Phenotyping

knowledge of the minor blood groups, their medical Table 1: Antigen frequencies of blood group antigens
significance and the laboratory methods such as typing Blood group Males Females n Frequencies (%)
and cross‑matching, etc.[7] A 111 43 154 24.64
B 140 77 217 34.72
Subjects and Methods AB 33 25 58 9.28
After taking oral informed consent, a total number of O 141 55 196 31.36
625 blood samples were studied, including 425 males Kell (K) 11 14 25 4
Duffy (Fya) 244 120 364 58.24
and 200 females. Blood donors were between 25
Duffy (Fyb) 156 93 249 39.84
and 29 years old (P  <  0.05) from both genders (male,
Kidd (Jka) 267 141 408 65.28
68%; female, 32%). Blood samples of both volunteer
Kidd (Jkb) 157 107 264 42.24
and replacement donors were collected into citrate MNSs (M) 360 183 543 86.88
EDTA (Ethylenediaminetetraacetic acid) vials. Extended N 188 101 289 46.24
antigen typing was performed for the major (ABO S 188 121 309 49.44
and Rh) and minor blood groups antigens including s 99 91 190 30.4
Kell, Duffy, Kidd, and MNSs by tube method using
commercially available antisera (Bio‑Rad Laboratories,
DiaMed, Switzerland). The statistical analysis was done Table 2: Distribution of genotype and phenotype
using Statistical Package for Social Sciences, version frequencies of blood group systems Duffy, Kidd, and
20.0 (IBM Corp., Armonk, NY, USA). Qualitative or MNS
categorical variables were described as frequencies and Genotype and allele Deduced phenotypesFrequency (%) n
proportions. Hardy–Weinberg equation was used to FY*A/FY*A Fya+Fyb‑ 34.56 216
calculate the gene frequencies where P  + q = 1 and P  = FY*A/FY*B Fya+Fyb+ 27.04 169
{2 × obs (AA) + obs (Aa)}/2 × {obs (AA) + obs (Aa) + FY*B/FY*B Fya‑Fyb+ 15.36 96
obs (aa)}; hence q = 1– p. JK*A/JKA* Jka+Jkb‑ 40 250
JK*A/JK*B Jka+Jkb+ 27.52 172
Results JK*B/JK*B Jka‑Jkb+ 13.6 85
M/M M+N‑ 53.92 337
The present study was aimed to determine the frequency M/N M+N+ 32 200
and differential expression of major and minor blood N/N M‑N+ 14.08 88
groups (Duffy, Kell, Kidd, and MNSs) antigens in
blood donors visiting blood bank of Shaheed Zulfiqar
Ali Bhutto Medical University (PIMS), Islamabad.
The distribution of antigen frequencies is illustrated in
Table 1. Most common phenotypes in Duffy, Kidd, and
MNSs blood group system were Fya + b‑, Jka + b‑, and
M + N‑, respectively [Table 2].
Gene frequencies
The calculated gene frequencies for minor blood
group systems were Kell K (0.04), Duffy Fya (0.481),
Fyb (0.519), Kidd Jka (0.538), Jkb (0.462), M (0.699),
N (0.301), S (0.445), and s (0.556) [Figure 1].
Figure 1: Gene frequencies of minor blood group antigens
Discussion
In humans, different combinations of blood group alloimmunization in multi‑transfused patients, young
antigens are present, producing a unique spectrum for females, and pregnant women. However, there is no
every individual. Distribution of blood group antigens such study until now on Pakistani population. In this
varies significantly in different populations and racial study, we examined the phenotype frequencies as well
groups. It is important to study the prevalence of as the genotype frequencies of clinically significant
different blood group antigens (both major and minor) blood group antigens in population of capital territory
not only for transfusion medicine but also for genetic Islamabad. The results of our study are compared
research and organ transplantation.[8] All the blood with other published data from neighboring countries
banks should have the data bank on various RBC including China, Iran, India, and Malaysia [Tables 3
antigens in local population to prevent the risk of and 4].

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Jabin, et al.: Red Blood Cells Phenotyping

The antigens of ABO blood group are found in the entire Duffy system, it has been reported previously that the
human population, but a considerable difference has Fya is very common among Asian populations. Similar
been observed in their frequency distribution all over results have been observed in this study, the incidence
the world.[13] Studies show that distribution of blood of Fya and Fyb antigen was 58.24% and 39.84%,
groups in different provinces of Pakistan considerably respectively, which is less as compared to India (87.3%
varies.[14] The current study has charted out that the and 58.3%).[5] The most prevalent phenotype in Duffy
distributional pattern of ABO blood group antigens as blood group was found to be Fya + Fyb‑ with 34.56%
follows: B > O > A > AB. Similar results were found which is significantly different to that reported in Iran,
out by previous studies in Pakistani population in both 22.8% and 47.4%,[10,18] India 41.7%,[5] 74% in Malays,
the genders.[15] With the Rh system, the present study 85.4% in Chinese, and 40.8% in Indians,[11] whereas
indicated 92% RhD+ and only 8% RhD‑ donors. These Caucasians and Africans have low prevalence of this
indications are consistent with the previous findings phenotype 17% and 9%, respectively.[12] In Kidd blood
among Pakistani population.[16] group system, Jk antigen acts as the urea transporter
The antigens of Kell blood group system are the third and its antibodies are difficult to detect. The antigen
most potent for triggering immune reaction.[3] The frequency of JKa and Jkb was recorded as 65.28% and
Kell (K) antigen was observed in 25 (4%) studied blood 42.24%, while Jka + Jkb‑ phenotype with the frequency
donors and its genotype was recorded as 0.04. Overall, of 40% was found to be the most common phenotype
there was a low prevalence of KK antigen among Asians which is found to be different with Malays 36%, Chinese
in China it was recorded 0,[9] and in the state of Parana, 24.5%, and Indians 35%.[11] At the same time, Persians
Southern Brazil, it was 0.0025 and 0.8.[4,17] With the showed the expression of this phenotype 34.7%,[10]
Caucasians 28% and Africans 57%.[12] In the MNS
Table 3: Comparison of genotype frequencies for the blood group system, the most common phenotype in our
Kell, Duffy, and Kidd blood group systems with other study was M+N-, with the frequency of 53.92% which
published data is much greater than observed in Indians (38.5%),[19]
Genotype Genetic frequency Genetic Genetic frequency Malaysians (37.5%), Chinese (34.3%),[11] Persians
(Islamabad) frequency (Parana) (n=400)[4] (43.3%),[10] Caucasians (28%), and 26% in Africans.[12]
(n=625) (China)[9] In case of S + s‑ phenotype, the prevalence was found
(n=146)
36.96%, which was about three times more common than
KK 0.04 0 0.0025
Fya 0.481 0.94 0.13
Indians 12.4%,[5] Persians 15,5%,[10] Caucasians 11%,
Fyb 0.519 0.06 0.39 Africans 3%,[12] Malays 1.5%, and Chinese 0.7%.[11]
Jka 0.538 0.51 0.27 These findings suggest a unique distribution pattern of
Jkb 0.462 0.49 0.25 some blood groups among the Pakistani blood donors.

Table 4: Comparison of most prevalent phenotype frequencies in the present study and other ethnic populations
Phenotype Current study (%) Study in Iran Study in Malaysia (Musa et al.)[11] Caucasians (Beadling Africans[12]
(Keramati)[10] (%) Malays Chinese Indians and Cooling)[12]
Duffy
Fya+Fyb‑ 34.56 47.4 74 85.4 40.8 17 9
Fya+Fyb+ 27.04 26.4 23 12.8 45 49 1
Fya‑Fyb+ 15.36 22.8 2.5 1.5 14.2 34 22
Fya‑Fyb‑ 23.04 3.4 0.5 0.4 0 v.rare 68
Kidd
Jka+Jkb‑ 40 34.7 36 24.5 35 28 57
Jka+Jkb+ 27.52 44.4 43 50.7 43.3 49 34
Jka‑Jkb+ 13.6 20.7 17.5 24.8 20 23 9
Jka‑Jkb‑ 18.88 0.2 3.5 0 1.7 v.rare v.rare
MNSs
M+N− 53.92 43.3 37.5 34.3 44.2 28 26
M‑N+ 14.08 13 18.5 22.6 18.3 22 30
M+N+ 32 43.7 44 43.1 37.5 50 44
S+s− 36.96 15.5 1.5 0.7 11.7 11 3
S+s+ 15.04 41.2 15.5 7.3 41.7 44 28

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Jabin, et al.: Red Blood Cells Phenotyping
The information about the minor blood group antigens
prevalence can assist in the management and prevention
of erythroblastosis fetalis and providing antigen‑negative
compatible blood to patients with multiple alloantibodies.
Based on the current study, it is inferred that a large
population carry the studied minor blood groups antigens
which can cause severe hemolytic transfusion reactions.
Conclusion
Thus, there is a need to perform the extended antigen
typing for minor blood groups as well, in all the blood
banks of Pakistan on a routine basis. This study provides
us a population profile with known antigens which can
be helpful to provide antigen‑matched blood to anemic
patients, pregnant women, and chronically transfused
patients.
Acknowledgment
The authors would like to thank the technical staff of
the blood bank at SZAB Medical University, Islamabad,
for providing assistance during research work and
Sindh Medical Stores, Lahore, for the availability of
antisera (Bio‑Rad Laboratories, DiaMed Switzerland).
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
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