1Department of Pharmacology, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia.
2Faculty of Medicine, Department of Hematology, King Abdulaziz University Hospital, Jeddah, Saudi Arabia.
3King Fahd Medical Research, King Abdulaziz University, Jeddah, Saudi Arabia.
4Center of Excellence in Genomic Medicine, KFMRC, King Abdulaziz University, Jeddah, Saudi Arabia.
Characterization of the ABO blood group at the phenotype and genotype levels is clinically essential for transfusion, forensics, and population studies. This study elucidated ABO phenotypes and genotypes, and performed an evaluation of their distribution in individuals from the western region of Saudi Arabia.
One-hundred and seven samples underwent standard serological techniques for ABO blood group phenotype analysis.
A phenotype distribution of 37.4%, 30.8%, 24.3%, and 7.5% was found for blood groups O, A, B, and AB respectively in our study cohort. Genotype analysis identified 10 genotype combinations with the
The study has a significant implication on the management of blood bank and transfusion services in Saudi Arabian patients.
This prospective study was approved by the ethical committee of the Faculty of Medicine, King Abdulaziz University Hospital at Jeddah (Saudi Arabia). Peripheral blood was obtained with informed consent from 107 participants. ABO phenotypes were serologically determined with anti-A and anti-B antibodies using an AutoVue Innova (Ortho Clinical Diagnostics, Raritan, NJ, USA).
Genomic deoxyribonucleic acid (DNA) from blood was extracted using the QIAamp DNA kit (Qiagen, Hilden, Germany). Briefly, 200 µL of sample was mixed with 20 µL of Qiagen protease in a microcentrifuge tube; 200 µL of buffer AL was added followed by thorough mixing and incubation at 56℃ for 10 min. Afterwards, 200 µL of absolute ethanol was added to the lysate, followed by gentle vortexing for proper mixing. Next, the mixture was transferred to a mini spin column for centrifugation and washed three time using buffer AWI before a final elution with 50 µL of AE buffer. Extracted DNA was quantified using a NanoDrop spectrophotometer (Thermo Fisher Scientific Inc., Wilmington, DE, USA). The method followed the procedure used by Muro et al. .
Primers used for PCR are listed in Table 1. The genotyping approach for this study was used previously in several studies [17, 18]. Four separate PCR reactions were performed using different sets of primers in each sample. Each 20 µL reaction contained 2 µL DNA mixed with 1 µL of each allele-specific primer (10 pmol), 10 µL master mix (2X), and 7 µL nuclease free water. PCR was performed in a thermal cycler (Applied Biosystems, Wilmington, DE, USA). The cycling conditions followed an initial denaturation at 95℃ for 3 min, followed by 35 cycles of denaturation at 95℃ for 40 s, annealing at 56℃ for 40 s, and elongation at 72℃ for 40 s, followed with a final elongation at 72℃ for 5 min. PCR products were separated by electrophoresis on 3% agarose gels containing ethidium bromide at 40 mV for 20 min followed by 70 mV for 30–40 min. A 100 base pair (bp) DNA ladder was used as a molecular marker (Invitrogen, Wilmington, DE, USA). The gel was visualized using a G-BOX UV transilluminator (Syngene, Cambridge, UK).
We found that serological
Out of 21 possible
Primers used to amplify the six alleles of
We found in our cohort from the western region of Saudi Arabia phenotype frequencies of 30.8%, 24.3%, 7.5%, and 37.4% for the A, B, AB, and O blood groups, respectively. Previous studies performed in other Saudi populations found different frequencies of 34.2%, 24.8%, 2.5%, and 38.5% for the A, B, AB, and O blood groups, respectively . For all studies including the present one, blood group O has the highest frequency followed by A, B, and AB. This variation in ABO phenotype found between different Saudi populations is expected among ethnic groups . For example, in the United Kingdom, blood group B has a low frequency (10%), whereas in India it is 18.8% .
We found 10
Of the six possible alleles, we found five alleles of which the most frequent allele was
As anticipated, the frequency of various alleles in different populations around the world can differ widely because of ethnicity (demonstrated in Table 4). This variation has been widely observed in many studies [5, 6, 8, 21, 22, 23, 24]. In the majority of populations examined, the
In conclusion, the present study reports the distribution of ABO phenotypes and genotypes in a group of individuals from the western region of Saudi Arabia. Using allele-specific multiplex PCR, 10
Electrophoretic pattern of the 10 identified