doi: 10.1111/iji.12114

Phenotypic and allelic distribution of the ABO and Rhesus (D) blood groups in the Cameroonian population S. T. Ndoula*,†,‡,1, J. J. N. Noubiap*,§,1, J. R. N. Nansseu*,¶ & A. Wonkam*,**

Summary Data on blood group phenotypes are important for blood transfusion programs, for disease association and population genetics studies. This study aimed at reporting the phenotypic and allelic distribution of ABO and Rhesus (Rh) groups in various ethnolinguistic groups in the Cameroonians. We obtained ABO and Rhesus blood groups and self-identified ethnicity from 14 546 Cameroonian students. Ethnicity was classified in seven major ethnolinguistic groups: Afro-Asiatic, Nilo-Saharan, Niger-Kordofanian/West Atlantic, Niger-Kordofanian/Adamawa-Ubangui, Niger-Kordofanian/Benue-Congo/Bantu/Grassfield, Niger-Kordofanian/Benue-Congo/Bantu/Mbam and Niger-Kordofanian/Benue-Congo/Bantu/Equatorial. ABO allelic frequencies were determined using the Bernstein method. Differences in phenotypic distribution of blood groups were assessed using the chi-square test; a P value 0.05). The frequencies of O, A, and B blood phenotypes were significantly lower, respectively, in the

* Faculty of Medicine and Biomedical Sciences, University of  I, Yaounde , Cameroon, † Guidiguis District Hospital, GuidiYaounde guis, Cameroon, ‡ Medical and Social Welfare Center of the Univera sity of Maroua, Maroua, Cameroon, § Internal Medicine Unit, Ede a, Cameroon, ¶ Mother and Child Centre, Regional Hospital, Ede , Cameroon and ** Division of Chantal Biya Foundation, Yaounde Human Genetics, Department of Clinical Laboratory Sciences, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa Received 11 October 2013; revised 20 December 2013; accepted 26 January 2014 Correspondence: Jean Jacques N. Noubiap, Internal Medicine Unit, a Regional Hospital, PO Box 100 Ede a, Cameroon. Ede Tel: 00237 79644081; E-mail:[email protected] 1

These authors contributed equally to this work.

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Nilo-Saharan group (P = 0.009), the Niger-Kordofanian/Benue-Congo/Bantu groups (P = 0.021) and the Niger-Kordofanian/West-Atlantic group. AB blood group was most frequent in the Niger-Kordofanian/ Adamawa-Ubangui group (P = 0.024). Our study provides the first data on ethnic distribution of ABO and Rhesus blood groups in the Cameroonian population and suggests that its general profile is similar to those of several sub-Saharan African populations. We found some significant differences in phenotypic distribution amongst major ethnolinguistic groups. These data may be important for blood donor recruitment policy and blood transfusion service in Cameroon.

Background ABO and Rhesus (Rh) are globally recognized as the most important of the 29 blood group systems. They are of critical importance in blood transfusion and underlie the occurrence of the haemolytic disease of the newborn. ABO and Rhesus (Rh) blood group systems are also of interest in population genetics studies, medicolegal investigations (Salaru, 1993; Segurel et al., 2013) and diseases susceptibility association studies (Miao et al., 2013; Pelzer et al., 2013; Lee et al., 2012). The frequencies of ABO and Rhesus (Rh) groups vary with geographical settings and populations ethnicity worldwide (Mourant et al., 1976). Cameroon is frequently referred to as ‘Africa in miniature’, because of its central location on the continent, its many geographical and cultural attributes and the great diversity of its population. Anthropological as well as population genetics studies have allowed the classification of Cameroonian population between three of the four major ethnolinguistic phyla found in Africa: the Afro-Asiatic phylum, the Nilo-Saharan phylum and the Niger-Kordofanian phylum (Tishkoff et al., 2009; Spedini et al., 1999; Bitjaa Kody, 2003). To the best of our knowledge and surprisingly, there is no published data on the ethnic distribution of ABO and Rhesus (Rh) groups in the Cameroonian population. Because blood group frequency figures are valid only for the specific population from which they are derived (Mourant et al., 1976), such data are therefore

© 2014 John Wiley & Sons Ltd International Journal of Immunogenetics, 2014, 41, 206–210

ABO and Rhesus blood groups in the Cameroonian population

needed in Cameroon. This study is aimed at reporting the phenotypic and allelic distribution of ABO and Rhesus (Rh) types in various ethnic groups in the Cameroonian population. It is anticipated that the data may contribute to strategize the supply and demand of bloods products in transfusion services nationwide and could have implications in investigating susceptibility to various conditions known to be associated with blood groups.

Data analysis

Data were coded, entered and analysed using the Statistical Package for Social Science (SPSS) version 20.0 for Windows (SPSS, Chicago, IL, USA). Descriptive statistics and parametric test (chi-square test) were used for comparisons. Significance level was set at 5%. Allelic frequencies were determined using the Bernstein method as follows (Nam & Gart, 1976): If p, q and r are, respectively, the allelic frequencies of allele A, B and O,

Methods Ethical considerations

Prior to commencing the study, a hospital local institutional ethical clearance was obtained. The study was performed in accordance with the guidelines of the Helsinki Declaration, and written informed consent was obtained from all the participants. Study population and setting

We conducted an observational cross-sectional study between October 2012 and April 2013 at the Medical and Social Welfare Center of the University of Maroua, a state that enrolled students from all over the country; the ethnic diversity of the Cameroonian population is anticipated to be represented in this student population. We consecutively included university students of Cameroonian parentage visiting the Medical and Social Welfare Center of the University of Maroua for a systematic medical evaluation who consented to participate in the study. Data collection

All participants provided demographic information including sex and self-identified ethnic affiliation. Ethnicity was classified in seven major ethnolinguistic groups: Afro-Asiatic, Nilo-Saharan, NigerKordofanian/West Atlantic, Niger-Kordofanian/Adamawa-Ubangui, Niger-Kordofanian/Benue-Congo/Bantu/ Grassfield, Niger-Kordofanian/Benue-Congo/Bantu/ Mbam and Niger-Kordofanian/Benue-Congo/Bantu/ Equatorial (Annex 1) (Tishkoff et al., 2009; Spedini et al., 1999; Bitjaa Kody, 2003; Ethnologue; Joshua project). We excluded subjects whose ethnic groups were unknown or difficult to classify. ABO and Rh blood group determination was carried out by Beth-Vincent and Simonin-Michon methods, which were performed on a slide, using commercial monoclonal blood grouping antisera: anti-A, anti-B, anti-AB and anti-D (Cypress Diagnostics, Belgium) and home-made red cell tests. We used a 3–5% washed red blood cell suspension that was prepared following a standard operating procedure described elsewhere (New Foundland Labrador, 2012).

© 2014 John Wiley & Sons Ltd International Journal of Immunogenetics, 2014, 41, 206–210

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p¼1

qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi freqðBÞ þ freqðOÞ

q¼1

qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi freqðAÞ þ freqðOÞ

r¼1

pffiffiffiffiffiffiffiffiffi freqðOÞ

Then, if p + q + r 6¼ 1, a correction must be carried out by the deviation D = 1  (p + q + r). So, the adjusted allelic frequencies are: p0 ¼ pð1 þ

D Þ 2

q0 ¼ qð1 þ

D Þ 2

r0 ¼ ðr þ

D D Þð1 þ Þ 2 2

The Hardy–Weinberg equilibrium in different ethnolinguistic groups was assessed by the goodness-of-fit chi-square test using the S2 ABOestimator software version 1.1.0.2 (Pedro J.N. Silva, Universidade de Lisboa, Portugal). The test is based on the following formula, where O represents the observed frequencies and E the expected ones (Schwartz, 1986): v2 ¼

P ðO  EÞ2 E

Results We analysed data from 14,546 subjects, 9759 (67.09%) being male. The ethnolinguistic groups Afro-Asiatic, Nilo-Saharan, Niger-Kordofanian/West Atlantic, NigerKordofanian/Adamawa-Ubangui, Niger-Kordofanian/ Benue-Congo/Bantu/Grassfield, Niger-Kordofanian/ Benue-Congo/Bantu/Mbam and Niger-Kordofanian/ Benue-Congo/Bantu/Equatorial represented, respectively, 28.65%, 2.17%, 4.61%, 23.90%, 22.45%, 3.24% and 14.98% of the studied population. In the ABO system, group O was the most frequent phenotype (48.62%), followed by A (25.07%), B (21.86%) and AB (4.45%) (Table 1). The distribution of ABO phenotypes in the studied population in total and amongst different ethnolinguistic groups did not

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Table 1. Distribution of ABO phenotypes in ethnolinguistic groups ABO phenotypes N (%) Ethnolinguistic groups

O

Afro-Asiatic Nilo-Saharan N-K/West Atlantic N-K/Adamawa-Ubangui N-K/B-C/Bantu/Grassfield N-K/B-C/Bantu/Mbam N-K/B-C/Bantu/Equatorial Total v2; P value**

2007 137 339 1633 1661 218 1077 7072 16.86;

A (48.16) (43.35) (50.52) (46.97) (50.87) (46.28) (49.43) (48.62) 0.009

B

1110 82 185 875 768 112 514 3646 14.96;

(26.64) (25.95) (27.57) (25.17) (23.52) (23.78) (23.59) (25.07) 0.021

873 82 124 776 708 121 496 3180 15.17;

AB (20.95) (25.95) (18.48) (22.32) (21.68) (25.69) (22.76) (21.86) 0.019

177 15 23 193 128 20 92 648 14.47;

(4.25) (4.75) (3.43) (5.55) (3.92) (4.25) (4.22) (4.45) 0.024

Total

Goodness-of-fit test of H-W frequencies v2; P value*

4167 316 671 3477 3265 471 2179 14546

1.3213; 0.6822; 0.6590; 2.5514; 0.2766; 0.7183; 0.1734; 0.3866;

0.2504 0.4088 0.4169 0.1102 0.5989 0.3967 0.6771 0.5341

*Degree of freedom: 1; **Degree of freedom: 6. H-W, Hardy–Weinberg; N-K, Niger–Kordofanian; B-C, Benue–Congo.

differ significantly from those expected under the Hardy–Weinberg law (P > 0.05). ABO allelic frequencies in ethnolinguistic groups are presented in Table 2. The Rh-positive blood group comprised 96.32% of the overall study population and Rh-negative 3.68% (Table 3). Blood group O Rhesus-positive represented about half of the entire population, while AB Rhesusnegative, found in approximately 1 over 1000 individuals, was the least frequent (Table 4). The frequencies

Table 4. Phenotypic distribution of ABO and Rhesus antigens Rhesus phenotypes ABO phenotypes

Rhesus +

Rhesus 

O A B AB

6797 3519 3074 620

275 127 106 28

(46.73) (24.19) (21.13) (4.26)

(1.89) (0.87) (0.73) (0.19)

Table 2. ABO allelic frequencies in ethnolinguistic groups Allelic frequencies estimates Ethnolinguistic groups

O

A

B

Afro-Asiatic Nilo-Saharan N-K/West Atlantic N-K/Adamawa-Ubangui N-K/B-C/Bantu/Grassfield N-K/B-C/Bantu/Mbam N-K/B-C/Bantu/Equatorial Entire sample

0.695893 0.663872 0.713855 0.682247 0.714178 0.684611 0.703969 0.697816

0.168837 0.168059 0.169632 0.167325 0.148274 0.152008 0.150441 0.160529

0.135269 0.168059 0.116509 0.150425 0.137548 0.163374 0.145590 0.141655

N-K, Niger–Kordofanian; B-C, Benue–Congo.

of O, A and B blood phenotypes were significantly lower, respectively, in the Nilo-Saharan group (P = 0.009), the Niger-Kordofanian/Benue-Congo/ Bantu groups (P = 0.021) and the Niger-Kordofanian/ West Atlantic group (P = 0.019). AB blood group was most frequent in the Niger-Kordofanian/AdamawaUbangui group (P = 0.024) (Table 1). We observed no significant difference in the frequency distribution of the Rhesus phenotypes between ethnolinguistic groups (Table 3). Table 5 shows the phenotypic distribution of ABO and Rhesus antigens in populations from different ancestries.

Discussion Table 3. Phenotypic distribution of Rhesus antigens in ethnolinguistic groups Phenotypic frequencies Ethnolinguistic groups Afro-Asiatic Nilo-Saharan N-K/West Atlantic N-K/Adamawa-Ubangui N-K/B-C/Bantu/Grassfield N-K/B-C/Bantu/Mbam N-K/B-C/Bantu/Equatorial Total v2; P value*

Rhesus + 4013 303 640 3343 3155 457 2099 14010 3.6906;

(96.30) (95.89) (95.38) (96.15) (96.63) (97.03) (96.33) (96.32) 0.7185

*Degree of freedom: 6. N-K, Niger-Kordofanian; B-C, Benue-Congo.

Rhesus 

Total

154 13 31 134 110 14 80 536

4167 316 671 3477 3265 471 2179 14546

(3.70) (4.11) (4.62) (3.85) (3.37) (2.97) (3.67) (3.68)

This study is the first to provide information on the ethnic distribution of ABO and Rhesus (Rh) phenotypes and alleles in the Cameroonian population. This study could serve as a preliminary nationwide report, because we recruited participants from a state university receiving students from all over the country and respecting the Cameroonian population. Africa contains more than 2000 distinct ethnolinguistic groups representing nearly one-third of the world’s languages (Ethnologue). Except for a few isolates that show no clear relationship with other languages, these languages have been classified in families and subfamilies within for major ethnolinguistic phyla: Niger-Kordofanian (spoken across a broad region of Africa), Afro-Asiatic (spoken predominantly in Saharan, north-eastern and eastern Africa), Nilo-Saharan

© 2014 John Wiley & Sons Ltd International Journal of Immunogenetics, 2014, 41, 206–210

ABO and Rhesus blood groups in the Cameroonian population

Table 5. Comparison of phenotypic distribution of ABO and Rhesus antigens in different populations Phenotypic frequencies Populations

O

A

B

AB

Rh +

Rh 

Cameroon present study Nigeriaa Guineab Madagascarc Mauritaniad Indiae Pakistanf Palestineg Turkeyh US White non-Hispanici US Hispanici US Black non-Hispanici US Asiani

48.6 46.6 48.9 41.6 49.1 38.7 28.2 32.0 30.8 45.2 56.5 50.2 39.8

25.1 23.05 22.5 22.6 28.3 21.9 23.8 40.0 43.8 39.7 31.1 25.8 27.8

21.9 25.95 23.9 29.7 18.6 32.7 38.0 22.0 16.2 10.9 9.9 19.7 25.4

4.4 4.4 4.7 6.1 4.0 6.7 10 6.0 9.2 4.1 2.5 4.3 7.1

96.3 96.4 95.9 98.9 94.2 91.8 89.1 97.3 86.0 82.7 92.7 92.9 98.3

3.7 3.6 4.1 1.1 5.8 8.2 10.9 2.7 14.0 17.3 7.3 7.1 1.7

a Kulkarni et al. (1985), bLoua et al. (2007), cRandriamanantany et al. (2012), dHamed et al. (2012), eDas et al. (2001), fHammed et al. (2002), gSkaik & EL-Zyan (2006), hDilek et al. (2006), iGarratty et al. (2004).

(spoken predominantly in Sudanic, Saharan and eastern Africa) and Khoesan (languages containing click consonants, spoken by San in southern Africa and by Hadza and Sandawe in eastern Africa) (Tishkoff et al., 2009). The first three of these phyla are represented in the Cameroonian population (Tishkoff et al., 2009; Spedini et al., 1999; Bitjaa Kody, 2003). This ethnolinguistic classification underlies numerous studies on the genetic structure and history of Africans and African Americans (Tishkoff et al., 2009). To the best of our knowledge, the presented is also the first in Africa to describe the differential distribution of blood groups based on this ethnic classification and could have implication for blood transfusion service planning and provision beyond Cameroon. Indeed, we found a significant difference in the distribution of ABO blood groups amongst the different ethnolinguistic groups, with the frequencies of O, A and B blood phenotypes being significantly lower, respectively, in the Nilo-Saharan, the Niger-Kordofanian/Benue-Congo/Bantu and the Niger-Kordofanian/West Atlantic groups (Table 1). As the AB group is rare, the collection of AB blood units could be directed particularly toward people of the Niger-Kordofanian/Adamawa-Ubangui ethnicity who present a significant higher frequency of this blood group (Table 1). There was no significant difference between ethnolinguistic groups with respect to Rhesus phenotypes. The gene frequencies with respect to ABO in this study can be represented as O>A>B>AB. O Rhesuspositive blood group was highly predominant, representing about half of the entire population, while AB Rhesus-negative was very infrequent. The distribution of the blood groups A, B, O and AB varies across the world according to the population. The predominance of the O phenotype is common in the majority of sub-

© 2014 John Wiley & Sons Ltd International Journal of Immunogenetics, 2014, 41, 206–210

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Saharan African populations (Table 5), but there are some slight differences in the distribution of phenotypes A and B (Tagny et al., 2009a,b; Kulkarni et al., 1985; Loua et al., 2007; Randriamanantany et al., 2012; Hamed et al., 2012). On the contrary, A and B phenotypes are predominant in some populations in the world: for instance, B phenotype has its highest frequency in Northern India and neighbouring Central Asia, and its incidence diminishes both toward the west and the east, falling to single digit percentages in Spain (Blood Transfusion Division, United States Army Medical Research Laboratory, 1971; Das et al., 2001; Hammed et al., 2002). It is believed to have been entirely absent from Native American and Australian Aboriginal populations prior to the arrival of Europeans in those areas (Ember et al., 2010). Blood group A is associated with high frequencies in Europe, especially in Scandinavia and Central Europe, although its highest frequencies occur in some Australian Aboriginal populations and the Blackfoot Indians of Montana [27–29]. As described by the present study, in most of sub-Saharan African populations, up to 90–95% of people have the phenotype Rhesus-positive (Tagny et al., 2009a,b; Kulkarni et al., 1985; Loua et al., 2007; Randriamanantany et al., 2012; Hamed et al., 2012). This frequency declines toward North Africa and are considerably lower in Caucasians (Tagny et al., 2009a,b; Lefrere & Rouger, 2009; Wagner et al., 1995; Garratty et al., 2004). To improve blood supply in African countries, the WHO recommends that in all African countries, there should be a National Blood Transfusion Centre aiming at implementing a coordinated blood supply across the country (World Health Organization, 2007). In a previous study conducted in a hospital-based blood bank in Yaounde, Cameroon, significant differences were found between blood donors and recipients with respect to ABO and Rhesus phenotypes (Tagny et al., 2009a,b). As the present data are population based and capture the full diversity of the population, it could greatly contribute to achieve the goal of the WHO recommendations in Cameroon, specifically, to strategize donor recruitment policies to assure the availability at all the time of the low-frequency blood groups. For example, we found that some blood groups were more predominant in some ethnolinguistic groups.

Conclusion Our study suggests that the ABO and Rhesus phenotypic profile of the general Cameroonian population is similar to several sub-Saharan African populations. We found some significant differences in the distribution of ABO blood groups between the different ethnolinguistic groups selected from the three main phyla Afro-Asiatic, Niger-Kordofanian and Nilo-Saharan. These data may be important for blood donor recruitment policies in a nationally coordinated blood transfusion service in Cameroon.

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Competing interests The authors declare that they have no competing interests.

Authors’ contributions STN designed the study, collected data, contributed to data analysis and revised the manuscript. JJNN designed the study, analysed the data, drafted and revised the manuscript. JRNN and AW critically reviewed and revised the manuscript. All the authors approved the final version of the manuscript.

Acknowledgements We are grateful to all the university students who participated in this research and to Dr. Rosette Amelie Youmbi for her support.

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© 2014 John Wiley & Sons Ltd International Journal of Immunogenetics, 2014, 41, 206–210

Phenotypic and allelic distribution of the ABO and Rhesus (D) blood groups in the Cameroonian population.

Data on blood group phenotypes are important for blood transfusion programs, for disease association and population genetics studies. This study aimed...
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