Distribution of ABO and Rh (D) blood groups and alleles among students of AlFashir University, El Fasher, Sudan. .Yahia Dawoud El Die1, Mustafa AbdAllah Yahya2, Al hafez Musa Ismail3 .Department of Biological Sciences, Al Fashir University, El Fasher, Sudan ,3 ,2 ,1
Corresponding author: yahiaeldie0@gmail.com
اﻟسودان, اﻟفاشر,ﺗﻮﺯيع ﺍﻟﺰﻣﺮاﻟدﻣوية والﻟيل ت وسط طل ب جاﻣعة اﻟفاشر مستخلص البحث في مجموع ة م ن طل بRh وABO أجريت الدراسة الحالية تقييما لتوزيع مجموعات الدم تسسم اكتشسساف الونسسواع الظاهريسسة لمجمسسوعه السسدم بواسسسطة. جامعة الفاشر بولية شمال دارفور وكان الهسسدف مسن هسذه الدراسسسة هسو تحديسسد مسسدي تسواتر الونسسواع.أسلو ب الشريحة الكلسيكية أظهسرت.الرئيسسية و الليلت لمجموعسة مسن الطل ب السسوداونيين المقيميسن فسي إقليسم دارفسور )125 ( يلﻴها الﺰمﺮﺓ%41)160 هي الكﺜﺮ اونﺘﺸارا إذ كاونتO الونتائج أن الﺰمﺮﺓ وأظهر ونظ ام. AB (سس%5.9)23 فالﺰمﺮﺓسB (سس%21)82 ثم الﺰمﺮﺓA (%32.1 كم ا كان.(O> A> B > AB ) في العيونة الجم اليه اتج اه الونتش ارABO مجموعه الدم . الس لبية و اليجابي ة مم اثلRh و ك ان أونتش ار.O > A > B تكرار الليلت بالترتي بسس و يجسس ب.الشخاص ذوي العامل الريسي السال ب كاونوا قلة في فسسي هسسذه الدراسسسة الستقصسسائية تقديم الونصح لمثل هذه المجموعات ليكوونوا حذرين في حياتهم و ذلك لصسسعوبة الحصسسول علسسي .هذا الونوع من الدم . طل ب, جاﻣعة اﻟفاشر, دراسة, ﻣجﻣوعة اﻟدم, ABO, Rh :اﻟكلﻣا ت اﻟﻣفﺗاحية
ABSTRACT Current study evaluated the distribution of ABO and Rhesus (Rh) D blood groups among the student population of Al Fashir University, North Darfur State. The blood group phenotypes were detected by the classic slide method. The objective of this study was to determine the major ABO phenotypes' and alleles’ frequency among a Sudanese student population residing in Darfur region. The results showed that, the highest blood group was O with 160(41%) followed by A blood group having 125(32.1%), B blood group 82(21%) and the lowest was AB blood group 23(5.9%). The ABO blood group system in the total sample showed the trend of prevalence (O≥ A> B > AB). However, the allelic frequencies were in the order of O > A > B. The Rh positive and negative distribution trend was also similar. People having the rhesus negative blood are few in this study. These categories should be advised to lead a very cautious life as their type of blood is very difficult to obtain. .Keywords: ABO, Rh, blood groups, study, Al Fashir University, students
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INTRODUCTION The human blood groups, the ABO system was discovered by Landsteiner in 1901 (Waters, 1996). This discovery marked the beginning of safe blood transfusion. The ABO antigens, although most important in relation to transfusion, also have variable expression on most tissues and are important histocompatibility antigens (Waters, 1996). The first attempt to classify races by genetic traits used the ABO system of blood groups. All people belong to one of four blood groups (A, B, AB or O), depending on which alleles of the ABO gene they inherited. The three major alleles of this gene, A. B and O, are present in almost all populations of the world but in different proportions (Cavalli-Sforza, 2009). Closely associated with the ABO system of blood grouping, is the rhesus system. The term rhesus is applied to any of the more than 30 substances, called agglutinogens, found on the surface of red blood cells, and are called rhesus factors denoted by the letters Rh. The Rh factors composition is unknown, and were named by the American pathologist, Karl Landsteiner and Alexander Solomon Wiener, who discovered the first of them in the blood of the rhesus monkey in 1937 (Microsoft Encarta, 2009). About 85% of people have this antigen and are said to be rhesus positive (Rh+). The remaining 15% have no rhesus antigen and are said to be rhesus negative (Rh-). Rh+ people do not make rhesus antibodies whereas Rh- people are capable of making anti-rhesus antibodies, (Waugh and Grant, 2001). The Rh factor is often associated with hemolyticanemia in the new born. A Rh- mother carries no Rh antigen on her red blood cells, but has the capacity to produce antibodies. If she conceives a child fathered by Rh+ man, and the baby inherits the Rh antigens from him, the baby may also be Rh+ (Waugh and Grant, 2001). During pregnancy, the placenta protects the baby from the mother’s immune system, but at delivery a few fetal red blood cells may enter the maternal circulation. When that happens, the maternal system will recognize these red blood cells as foreign because they carry the Rh antigen. This will stimulate the mother’s immune system to produce neutralizing antibodies to it. For the first baby there may be no danger. The red cells of the second and subsequent Rh+ babies are attacked by the maternal antibodies, which can cross the placenta and enter the fetal circulation. In the most severe cases, the baby dies in the womb from profound anemia. In less serious circumstances, the baby is born with some degree of anemia, which can be corrected with blood transfusion (Waugh and Grant, 2001). Rh disease usually results in jaundice, anemia, brain damage and often death, either before or shortly after birth (Microsoft Encarta, 2009). To 2
avoid this complication, now most men and women are tested for Rhfactor before having children. In addition an Rh- woman is given an injection of a substance called Rhogam immediately after delivery. Rhogam is a gamma globulin containing antibodies against the Rhfactor. By killing any of the Rh+ red cells that have migrated into the mother’s circulation from fetus, Rhogam prevents the mother from building up immunity to Rh- factor. Thus, subsequent Rh+ children are not exposed to anti Rh- antibodies. This procedure has largely eliminated Rh disease (Microsoft Encarta, 2009). As with many other genetic traits, the distribution of ABO and Rh blood groups varies significantly between populations and countries. Other blood group systems have been identified reaching up to thirty two in number. The distribution of ABO and Rh blood groups has been repeatedly investigated in various populations all over the world during the last half-century. The frequencies exhibit considerable variation in different geographic locations, reflecting the underlying genetic and ethnic diversity of human populations (Cavalli-Sforza et al, 1994). Thus in additions to ABO and Rh antigens, many other antigens are expressed on the red blood cell surface membrane. For example, an individual can be AB, Rh+ and at the same time M and N positive (MNS system), K positive (Kell system), Lea or Leb negative (Lewis system), and so on, being positive or negative for each blood group system antigen. Many of the blood group systems were named after the patient in whom the corresponding antibodies were initially encountered. The objective of this study was to determine the major ABO phenotypes' and alleles’ frequency in a Sudanese student population residing in Darfur region.
MATERIALS AND METHODs This study was conducted during the period extending from October 2016 to March 2017. It was done in the department of biological sciences, Faculty of Education, Al fashir University, El fasher, Sudan. El fasher is the administrative headquarter and capital city of North Darfur State, Sudan. Al fashir University was established in 1990. It is located in western area of the city. It has a student population of about 15,000 (DAA, 2017). Majority of them came from the 18 local government areas of the state. Less than 10% of the students came from other states of Sudan. The volunteers were randomly selected from the student population irrespective of sex, age or year of admission. Their blood groups were determined based on the slide method using a clean white wall tile. Three drops of blood were obtained from each volunteer from the tip of middle ring finger of left hand pricked with a sterile lancet. 3
Surface of finger cleaned with an alcohol swab for sterilization. The blood samples were placed on three different locations on the tile. Then a drop of each blood serum was placed on the drop of blood. Three type of sera were employed, anti-A serum for determining blood group A, anti-B serum for blood group B and anti-D serum for rhesus factor determination. Agglutination or clumping of red blood cells in anti-A serum indicates blood group A, and clumping of cells in anti-B serum indicates blood B. No clumping or agglutination of cells in both anti-A and anti-B sera indicate blood group O. Agglutination of cell in both anti-A andanti-B sera indicates blood group AB. Agglutination of blood cells in any blood type in anti-D serum indicates rhesus positive (+) and no agglutination indicates rhesus negative (-). An authorization to carry out the study was obtained from a local ethics. All the information that was obtained about the students was kept confidential. Allele frequency were calculated under the assumption of Hardy- Weinberg equilibrium and expressed as percentages. The frequencies of ABO in the study population were compared with the frequency of ABO in some other countries.
RESULTS Study Population The study population consisted of 390 students (121males, 269 females). The percentage of males was 31% while that of females was 69% (figure .1) 300
269
250 200 No. Subjects
150
121
100 50
69 31
frequency percentage
0 Sex
Figure 1: Distribution of the study students according to gender (n = 390).
Phenotypic Frequency of ABO Blood Groups Blood grouping was done by antigen antibody agglutination test by using commercial monoclonal anti sera. The distribution of blood group 4
phenotypes in the total sample were 32.1% (125), 21% (82), 5.9% (23), 41% (160) for groups A, B, AB and O, respectively. 180
160
160 140
125
120 100 No. Subjects
82
80 60 40
32.1
20 0
41
Frequency Percent
23
21
5.9 A
B
AB
O
Phenotype
Figure 2: Distribution of ABO groups among study population (n = 390).
The Allele Frequencies of ABO Antigens . Allele frequency for the antigens was computed by the Hardy-Weinberg law, on the basis of the number of students with blood groups, ABO. The distribution of the alleles in the total samples was 0.21, 0.15, and 0.64 for IA, IB and IO, respectively. The observed genotypes as compared with the phenotypes are shown in (Fig .3). The percentages of the alleles in our student sample population were : O+ and O- alleles : 0.31 and 0.10, respectively , while A+ : 0.28 , A-: 0.038 , B+ : 0.182, B-: 0.028, AB+: 0.052, AB- : 0.008. The Rh+ blood is the dominant blood in this survey and account for 83.5%. Blood group O+ is the predominant blood group (31%) followed by A+ (28.2%), B+ (18.2%), O- (10%), AB+ (5.2%), A-(3.8), B- (2.8%) and AB- (0.8). Plus or minus attached to blood groups indicate Rhesus positive and Rhesus negative respectively. People having the rhesus negative blood are few in this study.
5
140 120
121 110
100 80 NO. OF SUBJECTS
71
60 40 20 0
31
28.2 15 3.8
18.2
A-
B+
A+
11 2.8 B-
20 5.2
30.8
AB+
AB-
39
frequency Percent
10 O+
O-
GENOTYPE
Figure 3: the distribution and percentage of ABO and Rh blood groups for Al Fashir University students. 350
326
300 250 200 No. Subjects
150 100
frequency Percentage
83.5
64
50 0
16.5 Rh+
RhRhesus Factor
Figure 4: Distribution and percentage of Rh blood groups among the study population.
DICUSSION The current study aimed to estimate the distribution of ABO and Rh blood groups among Al fashir University student population in North Darfur, Sudan. The study included of 390 healthy student samples (121 males, 269 females).The frequencies of ABO and Rh blood groups vary from one population to another. The knowledge of the distribution of ABO and Rh blood groups is essential for effective management of blood
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banks inventory, to a facility of a smaller local transfusion service or a regional or national transfusion service. Apart from their importance in blood transfusion practice, the ABO and Rh blood groups are useful in population genetic studies, researching population migration patterns, as well as resolving certain medico-legal issues, particularly of disputed parentage. Also, ABO genotypes are important evidence at crime scenes, and for personal identification in forensic investigations and paternity testing. The results of the current study revealed that the most frequent blood group was O followed by A blood group, B blood group and the lowest was AB blood group. These results are consistent with the findings of (Hamed et al, 2014), who reported that O group was most frequent, 178(43.9%) and AB 26(6.4%) was the less frequent, where B group 104(25.7%) rank before A group 97(23.9%) disagreeing with our results. This study results are in agreement with (Shahata et al. 2012), who reported that the frequency of ABO blood phenotypes were as follows: O (51.5%), A (29.5%), B (16%), and AB (6%). Also the current results agree with the findings of Omotade et al, 1999. Who reported that, blood group O was ranked first with 54.2%, A 21.6%, B 21.4%, and AB 2.8%. The results of this study showed that subjects having blood group Rh+ 326(83.5%) higher than Rh- 64(16.5%) (Fig.4). Our results in consistence with the findings of (Hamed et al, 2014) who reported that, frequency of positive Rh blood group recording 375(92.6%) higher than negative Rh with 30(7.4%). Also our findings are however, consistent with many other studies reported higher prevalence of Rh+ including (Shahata et al., 2012; Omotade et al, 1999 and Olubayode, et al, 2013). But their results were slightly higher than recorded in the current study. The frequency of the alleles in the total was 0.214 for I A allele, 0.145 for IB allele and 0.641 for IO allele. This distribution is in agreement with the distribution with samples reported in an Iranian study where the reported: 0.2397 for IA allele, 01815 for IB allele and 0.5788 for IO allele (Nojavan et al, 2012). The frequencies of IA, IB and IO alleles in our subjects are also comparable to those reported in Iraqi and Jordanian studies where they found 0.212 for IA allele, 0.177 for IBallele, and 0.661 for IO allele and 0.270 for IA allele, 0.130 for IB allele and 0.60 for IO allele, respectively (Tills et al, 1983; Irashaid et al, 2002). The results showed that the frequency of IO allele is higher than either I A or IB and that of IA is higher than IB i.e., the trend is IO> IA> IB. CONCLUSION AND RECOMMENDATION
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In conclusion it can be said that blood group O is the predominant blood group followed by A then B blood group among the students of Al fashir University, El fasher, Sudan. It is recommended especially to the Al fashir University authority to make it mandatory for students to test their blood group at point of admission. This will help in reducing this blood group illiteracy. Mass campaign for the awareness of this basic health parameter should be encouraged. People with the rhesus negative blood should be advised to lead a cautious life so as to avoid blood transfusion. This is because blood with negative Rhesus factor is very difficult to come by during grouping and cross matching.
Acknowledgements Authors are grateful to Al Fashir University and ministry of Health, North Darfur State, Sudan, for facilitating this work.
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