Dataset Information

HV2 fragment mutations in ?-thalassemia patients and a new base pair insertion of high-altitude cases.

ABSTRACT: Worldwide, thalassemia represents one of the most common genetic disorders. There is a prevalence of Beta-thalassemia in Kingdom of Saudi Arabia (KSA), however there is a genetic counseling availability and an existence of mandatory premarital testing policy. Few studies detect molecular mutations of thalassemia genes in different KSA governates, including Makkah, Hufuf, Qatif, and Dammam but in our peer knowledge there is no reports on high altitude Taif region. The aim of the present study is to evaluate the molecular mutation analysis of ?-thalassemia gene in El Taif province (as a high-altitude area) patients of KSA and to estimate the iron overload toxicity due to thalassemia syndrome on the hotspot noncoding D-loop region (hypervariable, HV2 gene fragment) of mtDNA. Blood samples were collected from total 25 ?-thalassemia patients and 25 normal control that were used for HPLC, hematological analysis and different molecular evaluations. Extracted nuclear DNA from blood sample was used to detect known mutations accompanied with ?-thalassemia in other countries using PCR-ARMS technique targeting IVSII-1, IVSI-5, Codon 8/9, Cd44 and Cd5 genes' mutations. Moreover, mtDNA was used to detect point of mutation of HV2 fragment in the D-loop region using PCR-SSCP and then sequencing. Results show significant increase in the level of HbA2 and decrease of HbA in comparison to control by using HPLC. PCR-ARMS reports that all ?-thalassemia patients have heterozygous alleles of wild and mutated regions with nucleotide transition/transversion of IVSI-5 (AC>AG), Codon 8/9 (CT>CC), and Cd44 (GG>GA), however no point of mutation was detected in IVSII-1 (AC>AT) Cd5 (CT>CG) genes. Moreover, PCR-SSCP shows points of mutations for ?-thalassemia HV2 fragment that were confirmed by sequencing in the form of base pairs deletion, insertion and transition/transversion. For the first time, the present study reports the presence of 2 bps found in HV2 region that might be specific to KSA nations and not found in other countries. In conclusion, our results were in concurrent with other studies in the presence of specific genetic mutations in ?-thalassemia patients that is accompanied with points of mutations in HV2 region of high altitude Taif governate.

SUBMITTER: Al Thwiby NM

PROVIDER: S-EPMC7675125 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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HV2 fragment mutations in β-thalassemia patients and a new base pair insertion of high-altitude cases.

Al Thwiby Nsreen M NM Hussien Nahed A NA Bayoumy Eman S ES Shokary Mohammed M

American journal of blood research 20201015 5

Worldwide, thalassemia represents one of the most common genetic disorders. There is a prevalence of Beta-thalassemia in Kingdom of Saudi Arabia (KSA), however there is a genetic counseling availability and an existence of mandatory premarital testing policy. Few studies detect molecular mutations of thalassemia genes in different KSA governates, including Makkah, Hufuf, Qatif, and Dammam but in our peer knowledge there is no reports on high altitude Taif region. The aim of the present study is ...[more]

PMID: 33224572

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Project description:BackgroundThe frequency of the beta-thalassemia (β-thalassemia) gene in Indonesia ranges from 3 to 10%. However, in the East Java province, there is still limited information on the prevalence of β-thalassemia mutations in clinically diagnosed beta-thalassemia patients of East Java. Therefore, this study aimed to characterize β-thalassemia mutations in selected patients in the East Java province of Indonesia.MethodsThis is an analytical observational study. Diagnosis of β-thalassemia was based on clinical presentation, complete blood count (CBC), and hemoglobin (Hb) electrophoresis. Blood specimens taken from each patient in three ethylenediaminetetraacetic acid (EDTA) tubes were analyzed for CBC and Hb electrophoresis and processed for DNA extraction and subsequent polymerase chain reaction (PCR). Detection of mutations in Hemoglobin Subunit Beta (HBB) gene exons 1-3 of the β-thalassemia gene as the common mutation in Indonesia was done using PCR followed by Sanger sequencing.ResultsIn total, 33 (n = 33) participants were involved in this study with ages ranging from 5 to 17 years comprising 19 women and 14 men. Their ethnic origins were Javanese (n = 30) and Chinese (n = 3). CBC results showed that mean ± standard deviation (SD) for Hb, red blood cell (RBC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and red cell distribution width (RDW)-CV were 81.2 ± 7.0 g/L; 3.40 ± 0.39 × 109/L; 71.05 ± 5.72 fL; 24.12 ± 2.45 pg; 33.91 ± 1.47 g/dl; 24.38 ± 6.02%, respectively. Hb electrophoresis revealed that 5 out of 33 participants had beta-thalassemia and 28 out of 33 participants had hemoglobinopathy (Hb) E/beta-thalassemia. Results of Sanger sequencing showed the following genotype variations in the samples: 12 (36.4%) with β CD26 /β IVS-I-5; 6 (18.2%) with β CD26 /β CD35; 3 (9.1%) with β CD26 /β IVS-I-2; 2 (6.1%) with β CD27/28 /β CD40; 2 (6.1%) with β IVS-I-1 /β CAP+1; and β CD26 /β IVS-I-1; β IVS-I-5 /β CAP+1; β IVS-I-5 /β CD35; β CD26 /β CD37; β CD26 /β CD15; β CD26 /β CD40; and β IVS-I-5 /β CD19 in 1 (3%) sample, respectively, and 1 (3%) had no abnormality detected in sequencing even though electrophoresis showed abnormality in the migration pattern. The β CD26 /β IVS-I-5 mutation was found in samples that were noted to have Hb E/beta-thalassemia on Hb electrophoresis.ConclusionThe underlying genetic variations are heterogeneous in thalassemia patients in East Java, where 12 variants were found. The most common variant was β CD26 /β IVS-I-5, which all accounted for Hb E/beta-thalassemia on Hb electrophoresis. Furthermore, 28 out of 33 participants had hemoglobinopathy (Hb) E/beta-thalassemia.

Dataset Information

HV2 fragment mutations in ?-thalassemia patients and a new base pair insertion of high-altitude cases.

Publications

HV2 fragment mutations in β-thalassemia patients and a new base pair insertion of high-altitude cases.

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