HbA2 Levels Research Articles

Evaluation of Haemoglobin F and Haemoglobin A2 among Sickle Cell Patients in Steady State at Selected Hospital in Ogun State, Nigeria

Background: Sickle cell anemia (SCA) is a genetic disorder characterized by abnormal hemoglobin variants, including hemoglobin F (Hb F) and hemoglobin A2 (Hb A2). Evaluating the levels of these hemoglobin variants in steady-state sickle cell patients can provide insights into disease prognosis and management. This study aimed to assess Hb F and Hb A2 levels among sickle cell patients in steady state at a selected hospital in Ogun State, Nigeria. Objective: To evaluate the levels of hemoglobin F (Hb F) and hemoglobin A2 (Hb A2) in sickle cell anemia patients and determine their associations with demographic factors such as age and gender. Materials and methods: A descriptive cross-sectional study was conducted at the Sickle Cell Center, Abeokuta, Ogun State, Nigeria, from September to October 2020. A total of 60 sickle cell patients in steady state were recruited using convenient sampling. Blood samples were collected, and Hb F and Hb A2 levels were measured using high-performance liquid chromatography (HPLC). Data were analyzed using SPSS version 26.0 with independent t-test and bivariate correlation. Results: The mean levels of Hb F and Hb A2 varied across age groups and genders. Hb F levels were highest in patients over 35 years (10.4 ± 12.16%) and lowest in those aged 26-30 years (3.7 ± 1.3%). The majority of participants had Hb F levels between 2-10%, with a significant association between Hb F levels and age (p = 0.038). Hb A2 levels were consistently above 3.1% in 92.6% of the study population, with no significant association with age or gender. Conclusion: There is a significant association between age and Hb F levels among steady-state sickle cell patients, while Hb A2 levels showed no significant demographic correlations. The findings suggest that Hb F may serve as a potential marker for clinical evaluation in SCA patients.

Association study of common KLF1 variants with Hb F and Hb A2 levels in β-thalassaemia carriers of Portuguese ancestry

Association study of common KLF1 variants with Hb F and Hb A2 levels in β-thalassaemia carriers of Portuguese ancestry

Analysis of five Chinese individuals with rare thalassemia mutation HBB: c.93-21G＞A

To explore the hematological phenotype and genotypic characteristics of five Chinese individuals with a rare thalassemia mutation HBB: c.93-21G＞A. A retrospective study was carried out on five individuals identified by the People's Hospital of Yangjiang and Guangzhou Hybribio Co., Ltd. from May 2018 to September 2022. Routine blood test and hemoglobin electrophoresis were performed, and the genotypes of five subjects were determined by using PCR combined with reverse dot blotting (RDB), nested PCR, Gap-PCR and Sanger sequencing. This study was approved by the People's Hospital of Yangjiang (Ethics No. 20240001). Among the five individuals, hematological data of one was unavailable, and the remaining four had presented with microcytosis and hypochromia. The results of hemoglobin electrophoresis indicated that all of them had a HbA2 level of ≥4.7%. Genetic analysis showed that one case had harbored compound heterozygous mutations of αααanti3.7 triplet and HBB: c.93-21G＞A, one had compound heterozygous mutations of -α3.7 and HBB: c.93-21G＞A, whilst the remaining three were heterozygous for the HBB: c.93-21G＞A mutation. The hematological phenotype of β-thalassemia carriers (HBB: c.93-21G＞A) is similar to that of other β+ thalassemia heterozygotes with mild β-thalassemia characteristics.

Loss-of-Function Variants in SUPT5H as Modifying Factors in Beta-Thalassemia.

It is well known that modifiers play a role in ameliorating or exacerbating disease phenotypes in patients and carriers of recessively inherited disorders such as sickle cell disease and thalassemia. Here, we give an overview of the literature concerning a recently described association in carriers of SUPT5H Loss-of-Function variants with a beta-thalassemia-like phenotype including the characteristic elevated levels of HbA2. That SUPT5H acts as modifier in beta-thalassemia carriers became evident from three reported cases in whom combined heterozygosity of SUPT5H and HBB gene variants was observed to resemble a mild beta-thalassemia intermedia phenotype. The different SUPT5H variants and hematologic parameters reported are collected and reviewed to provide insight into the possible effects on hematologic expression, as well as potential disease mechanisms in carriers and patients.

Coexistence of Iron Deficiency Anaemia (IDA) and Beta Thalassaemia Trait (B-TT)

Background: The WHO estimates that about 7% of the world population are thalassaemia trait and increasing number of cases are being detected. The world population of carrier of b thalassamia is reported to be more than 100 million. Bangladesh also lies in thalassaemic Belt. A conservative world health report estimates that 3% of our populations are carrier of b thalassaemia which means that there are 3-6 million of b thalassaemia are in Bangladesh. To manage thalassaemia and thalassaemia trait, our traditional concept is not to give iron or iron conaining food. But concomitant iron deficiency anaemia in thalassaemia trait patient may be fatal. Those patients should be evaluated by serum ferritin or by the iron profile and treatment should be supplemented with rational amount of iron. Methodology: This cross sectional observational study was carried out on 75 patients from 03 to 59 years of age of both sex of Beta Thalassaemia Trait, in the department of Medicine and Haematology of Dhaka Medical College Hospital (DMCH) and Bangabandhu Sheikh Mujib Medical University (BSMMU) from January, 2015 to December, 2015. The patients previously diagnosed as Beta Thalassaemia Trait was included in study population. Result: In this study out of 75 Beta Thalassaemia Trait patients, 21 had evidence of Iron Deficiency Anaemia (IDA) and 54 patiens had no evidence of IDA. Thus the frequency of coexistent Beta Thalassaemia Trait and IDA in this study is 28%. The mean Hb concentration was 9.66 gm/dL (±1.54) which is 8.48 gm/dL (±1.43) and 10.12 gm/dL(±1.34) in Beta Thalassaemia Trait with IDA patients and Beta Thalassaemia Trait without IDA patients respectively. So the Hb concentration is significantly lower in b-TT with IDA patients than those without IDA (p-value < 0.001). The mean of Hb A2 level was 4.87(%) (±0.54) which is 4.44(%) (±0.40) and 5.03(%) (±0.50) in Beta Thalassaemia Trait with IDA patients and Beta Thalassaemia Trait without IDA patients respectively. So the Hb A2 level is significantly lower in Beta Thalassaemia Trait with IDA patients than those without IDA (p-value < 0.001). Conclusion: From this study, it could be concluded that the frequency of IDA in Beta Thalassaemia Trait is about 28%. The degree of anaemia is more severe and the level of Hb A2 is much lower in Beta Thalassaemia Trait with IDA patients than Beta Thalassaemia Trait without IDA. J Dhaka Med Coll. 2022; 31(2) : 182-186

Distinguishing Iron Deficiency Anemia From Beta-Thalassemia Trait: Comparative Analysis of CRUISE Index and Other Traditional Diagnostic Indices.

Introduction Iron deficiency anemia and beta-thalassemia trait are two common and important differentials of microcytic hypochromic anemia. Various discrimination indices using two or more common complete blood cell count (CBC) parameters have been used to distinguish between the two since1973. Recently, a new discriminant index, the CRUISE index, was proposed in the year 2019. The efficacy of various older indices along with the CRUISE index was evaluated for patients in our geographical area. Materials and method Ours was a laboratory-based, cross-sectional study where 100 patients, based on inclusion and exclusion criteria, with microcytic hypochromic anemia were evaluated for CBC parameters along with serum ferritin and hemoglobin-high performance liquid chromatography (Hb HPLC). A total of eight discrimination indices namely, Mentzer, Srivastava, Shine & Lal, Green & King, RDWI, England & Fraser, Kerman I and CRUISE index were used and evaluated for their diagnostic efficacy using different statistical parameters. ROC curves were obtained and a new cut-off value was proposed for our population. Data was analysed using Microsoft Excel (Microsoft® Corp., Redmond, WA, USA) and SPSS v29.0.2.0 (20) (IBM Corp., Armonk, NY, USA). Results Out of the total 100 cases, 39 were beta-thalassemia trait and 61 were iron deficiency anemia cases. The average age was 36.7 (±12.7 SD) years. Among the 73 females, 43 were diagnosed as iron deficiency anemia (IDA) and 30 as beta-thalassemia trait (BTT) cases. Among the 27 males, 18 were diagnosed as IDA and nine as BTT cases. The mean values were significantly lower in IDA patients for mean corpuscular volume (MCV) (p=.008), mean corpuscular haemoglobin (MCH) (p=.003), and mean corpuscular haemoglobin concentration (MCHC) (p=.003) and significantly higher for red cell distribution width (RDW) (p=.020). The mean ferritin levels in cases of IDA were 7.61 (±3.75) mcg/L and in BTT were 87.09 (±66.77 SD) mcg/L. The mean HbA2 levels in IDA cases were 2.75% (±0.41% SD) and BTT cases were 5.57% (±0.73% SD). CRUISEindex revealed the highest AUC (0.934), YI (76.21) and accuracy (90%) followed by the Mentzer index with a diagnostic accuracy of 81%. Shine & Lal index revealed the lowest AUC (0.710), YI (3.28) and accuracy (41%). Conclusion CRUISE index, which was recently proposed, was ranked 1st in terms of AUC, YI, and accuracy and was considered 2nd best in terms of sensitivity for differentially diagnosing the two conditions. Mentzer index, a commonly used index, also revealed a high diagnostic accuracy in our study for differentiating BTT from IDA. CRUISE index being a novel index, more research work needs to be carried out in various other geographical setups to evaluate the efficacy of this index.

Serum Iron and Ferritin Levels in Beta Thalassemia Carriers in Duhok Governorate, Iraq.

Background Thalassemia and iron deficiency anemia (IDA) account for most cases of microcytic hypochromic anemia. It is a common misconception that iron deficiency does not occur in thalassemia. However, studies have found that iron deficiency can coexist in carriers of beta thalassemia. Objective The aim of this study was to determine the prevalence of iron deficiency and iron overload in carriers of beta thalassemia in Duhok, Iraq. Patients and methods This prospective cross-sectional study included 250 patients with beta thalassemia carriers attending Kurdistan Private Hospital Laboratory Department from July 2021 to June 2023. Patients with microcytic hypochromic blood picture were tested for HbA2 levels, and those with a level >3.7% were included in the study and were tested for serum iron and serum ferritin levels. Results The age range was 15-80 years, with a mean of 25 years, and the male-to-female ratio was 1.5:1. The prevalence of iron deficiency in beta thalassemia carriers was 16% (N = 40). The prevalence of iron overload was 8.4% (N = 21). There was a significant statistical difference among those with iron deficiency, normal iron status, and iron overload in terms of hemoglobin level (P=0.001), RBC count (P=0.012), HbA2 (P=0.015), and serum ferritin (P < 0.0001). Conclusion Iron deficiency is more prevalent in beta thalassemia carriers than iron overload, necessitating proper assessment of iron status in patients with beta thalassemia carriers. Those with abnormal iron status need effective treatment to optimize the overall outcomes in patients with beta thalassemia carriers.

Thalassemia Genotypes and HbA2 levels of Children in Wuzhou, Guangxi

To analyze thalassemia genotypes and distribution of children in Wuzhou Guangxi, and evaluate the diagnostic value of HbA2 in children's thalassemia screening, so as to provide scientific evidence for the prevention and control strategies of thalassemia. Four hundred and fifty-eight children suspected with thalassemia in Wuzhou were enrolled from March 2017 to June 2022. The level of HbA2 was detected using Bio-Rad VARIANT II Hb analysis system. The deletion of α-thalassemia was measured with gap-PCR assay, and the point mutation of α- and β-thalassemia was tested with DNA reverse dot blot hybridization assay. The receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of HbA2 for children's thalassemia. A total of 304 thalassemia carriers were detected in 458 children, accounting for 66.38%. One hundred and seventy-five cases were defined to be α-thalassemia, with the main type of --SEA/αα (54.86%). Thirty-six cases were defined to be intermediate α-thalassemia, with the main type of -α3.7/--SEA (9.72%). In 108 cases with β-thalassemia, βCD41-42/βN was the main type, accounting for 49.07%, followed by βIVS-Ⅱ-654 /βN (14.81%). Seven cases were moderate/severe β-thalassemia (predominantly β-28/β-28 and βCD41-42/βCD17/). Twenty-one genotypes of α- and β-thalassemia were found in the children. There was significant difference of HbA2 level between the children with different types of thalassemia and healthy controls (all P < 0.001). ROC curve analysis showed that the sensitivities of HbA2 for α-thalassemia, β-thalassemia and αβ-thalassemia were 74.3%, 82.4% and 85.7%, with the optimal cut-off values of 2.60%, 3.60% and 3.70%, respectively, the specificities were 64.3%, 96.1% and 96.8%, and the area under the curve were 0.690, 0.887 and 0.916, respectively. The thalassemia genotypes of children in Wuzhou are diverse. It is necessary to further strengthen the prevention and control measure of thalassemia to reduce birth defects and improve birth quality.

Exploring the Clinical and Hematological Characteristics of Beta-Thalassemia Trait: A Comprehensive Analysis in a Tertiary Care Hospital Setting.

Beta-thalassemia is one of the most common inherited hematological diseases caused by more than 350 mutations in the β-globingene (HBB). Beta-thalassemia carrier or trait is associated with defects in one allele of the HBB gene. The majority of beta-thalassemia trait cases remain concealed in society and remain unnoticed as they are mostly asymptomatic or present with mild symptoms of anemia. There is a 25% chance of having children with beta-thalassemia major and a 50% chance of having carrier babies when two people with beta-thalassemia trait are married. Hence, it is important to identify the individuals with beta-thalassemia trait and provide counseling to understand the risks of pregnancy and its outcome. Aim To study the identification of beta-thalassemia trait cases along with their clinical findings and hematological correlation. Materials and methods Study Design This was a retrospective study conducted at Saveetha Medical College and Hospitalfor a period of four years from January 2020 to December 2023. Inclusion Criteria Age group more than 18 years, antenatal mother, cases of anemia who were refractory to iron treatment, and screening of family members in the positive cases of beta-thalassemia trait. Exclusion Criteria History of blood transfusion within three months was excluded. Data Collection A total number of 837 cases were screened to rule out the presence of beta-thalassemia trait/hemoglobin (Hb) variants. A 2 mL of intravenous blood samples were collected in an ethylene diamine tetraacetic acid (EDTA) vacutainer tube and processed in a Sysmex XN 1000 (Hyogo, Japan: Sysmex Corporation)automated hematology analyzer. The hematological parameters were analyzed. Statistical Analysis The study included both descriptive and analytical characteristics. Mean and standard deviation (SD) were calculated for all the hematological parameters. Beta-thalassemia trait was diagnosed with an HbA2 level of more than 4.0% through high-performance liquid chromatography (HPLC) analysis. Results Among the 837 samples studied for HPLC screening, 74 (8.8%) cases were found to have beta-thalassemia trait. The age group included was from 18 years to 56 years. Of 74 cases studied, 32(43%) were females and 42(57%) were males. Among the 74 cases studied, the Mentzer index <13 was seen in 58 (78%) cases and the Mentzer index >13 was seen in 16 cases (22%). Thirty-four cases (46%) of beta-thalassemia traits presented to the hospital with a history of fever for evaluation and antenatal screening accounted for 19 cases (26%). The mean red blood cell (RBC) count was 5.5 million/cu.mm; mean corpuscular volume (MCV) was 63.8 fL; mean corpuscular hemoglobin (MCH) was 19.6 pg; red cell distribution width coefficient of variation (RDW-CV) was 17.8%. Among the 74 cases studied, 37(46%) cases had an Hb of more than 11 g/dL, 22 cases had mild anemia, 12 cases had moderate anemia, and three cases had severe anemia. Conclusion This study concluded that regular monitoring of the Mentzer index along with HPLC analysis is an effective approach in identifying beta-thalassemia trait cases and further providing genetic counseling among the couples that will help in reducing high-risk pregnancy and the birth of a child with thalassemia major.

Molecular characterization of similar Hb Lepore Boston-Washington in four Chinese families using third generation sequencing.

Hemoglobin (Hb) Lepore is a rare deletional δβ-thalassemia caused by the fusion between delta-beta genes, and cannot be identified by traditional thaltassemia gene testing technology. The aim of this study was to conduct molecular diagnosis and clinical analysis of Hb Lepore in four unrelated Chinese families using third generation sequencing. Decreased levels of mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and an abnormal Hb band were observed in the probands of the four families. However, no common α and β-thalassemia variants were detected in the enrolled families using polymerase chain reaction-reverse dot blot hybridization based traditional thalassemia gene testing. Further third-generation sequencing revealed similar Hb Lepore-Boston-Washington variants in all the patients, which were resulted from partial coverage of the HBB and HBD globin genes, leading to the formation of a delta-beta fusion gene. Specific gap-PCR and Sanger sequencing confirmed that all the patients carried a similar Hb Lepore-Boston-Washington heterozygote. In addition, decreased levels of MCH and Hb A2 were observed in the proband's wife of family 2, an extremely rare variant of Hb Nanchang (GGT > AGT) (HBA2:c.46G > A) was identified by third-generation sequencing and further confirmed by Sanger sequencing. This present study was the first to report the similar Hb Lepore-Boston-Washington in Chinese population. By combining the utilization of Hb capillary electrophoresis and third-generation sequencing, the screening and diagnosis of Hb Lepore can be effectively enhanced.

Red Cell Parameters in Beta Thalassemia Trait; Comparison between Iron Deficient and Non-Deficient Carriers

Objective: To compare hematological parameters in iron-deficient and non-deficient carriers of beta thalassemia trait. Study Design And Setting: Comparative cross-sectional study. This study was conducted at the Department of Hematology at PNS SHIFA Hospital, Karachi for six months (February 2023 to July 2023). Methodology: This comparative cross-sectional study included a total of 304 cases with beta thalassemia trait and was divided into two groups; the Iron-deficient (ID+) and iron-deficient (ID-) group. The study focused on red cell parameters i.e, Hemoglobin (Hb), TRBC, MCV, MCH and RDW. Haemoglobin Electrophoresis results of both groups were also compared. Descriptive statistics were expressed as mean ± SD and the Chi-square test was assigned. A p-value = 0.05 was considered statistically significant. Results: Out of the total 304 subjects, 76 (25.0%) had iron deficiency, and 228 (75.0) had sufficient iron stores. Mean age of patients was 18 (Range: 3 – 35) years. HbA levels were similar in both the groups with p-value &gt; 0.05. While all other parameters compared showed marked differences among the two groups and were found to be statistically significant. Hb, TRBC, MCV MCH showed lower values in the ID+ group while RDW was lower in the ID- group. Conclusion: Red Cell parameters and HBA2 levels in beta thalassemia trait vary significantly among the iron deficient vs non-deficient group. Hence due consideration is needed in screening of beta thalassemia trait in patients with iron deficiency anemia.

Role of blood hepcidin alteration as a biomarker in β-thalassemia patients’ diagnosis

The objective of this research was to determine the change in hepcidin levels and other biochemical markers, including total iron binding capacity (TIBC), serum iron, testosterone, and specific vitamins in the blood of individuals with β-Thalassemia. Here, 140 participants were involved in the study, of whom 110 were affected by β-thalassemia and 30 were healthy. The samples were obtained from Baqubah Teaching Hospital, the blood bank and blood donation center. The study was carried out from May 2022 to August 2022, and the patients were housed in Diyala Provence, Baquba City and its suburbs. The participants in this investigation were split into three groups: A: 55 male patients with β-thalassemia; B: 55 female patients with β-thalassemia; and C: 30 healthy individuals that were set as a control group. The findings of the study showed that the levels of HbA1 in males and females were 93.44 ± 0.71 and 93.53 ± 0.91, respectively, while the levels of HbA2 in males and females were 4.99 ± 0.59 and 5.29 ± 0.72, respectively. In contrast, the control group had HbA1 levels of 97.33 ± 0.40 in males and 97.66 ± 0.46 in females, but HbA2 levels were 2.32 ± 0.33 in males and 2.24 ± 0.24 in females. The study revealed remarkable differences (P &lt; 0.05) between these variables. Hematological measures, such as hemoglobin concentration and percentages of mean corpuscular volume (MCV), packed cell volume (PCV), and mean corpuscular hemoglobin (MCH) were substantially reduced (P &lt; 0.05) in β-thalassemia patients when compared to the controls. Serum iron and TIBC were significantly increased (P &lt; 0.05), while Hepcidin levels were markedly decreased (P &lt; 0.05) in the serum of β-thalassemia patients compared to controls. Therefore, as a conclusion the reduction of hepcidin levels and increase in iron levels are correlated with β-thalassemia and can be used as a biomarkers in monitoring β-thalassemia disease.

HBA2 levels in iron deficiency - Can iron deficiency mask thalassemia screening?

HBA2 levels in iron deficiency - Can iron deficiency mask thalassemia screening?

Anemia among Medical Students from Jakarta: Indonesia-Iron Deficiency or Carrier Thalassemia?

Anemia, a global health concern, affects one-fourth of the global population, particularly women. In Indonesia, its prevalence is 23.7%, with 32.0% among 15-24 year-olds. Factors include poor nutrition, infectious diseases, chronic diseases, inherited disorders, and inadequate healthcare access. This study aimed to investigate anemia prevalence and its etiology among medical students from Jakarta. This study was a descriptive research with a cross-sectional approach. Undergraduate students aged 18-23 years old were selected and consented to participate by a consecutive nonrandom sampling methods. Laboratory blood data were evaluated (including Hb, MCV, MCH, HbA2, and ferritin levels) and DNA was isolated to confirm the type of thalassemia carrier. In total, 140 medical students, mainly female, were recruited. Anemia was found in 13.6% (11.4% had low MCV and/or MCH), and 16.5% had low MCV and/or MCH without anemia. Hb electrophoresis revealed high HbA2 values, suggesting the HbE variant (2.1%), and β-thalassemia carrier (0.7%). DNA analysis confirmed the cd26 mutation and heterozygous IVS1nt5. Among those without anemia, 5% had α-deletion, while in the group with anemia, 1.4% had α-deletion (with coexistent IDA), 3.6% had α-deletion, and 0.7% had β-mutation. DNA analysis can identify specific mutations associated with alpha-thalassemia, distinguishing between iron deficiency anemia and the alpha-thalassemia trait. Thalassemia screening should involve low MCV and/or MCH values as the first step (stage 1), followed by Hb analysis (stage 2) and DNA analysis (stage 3). In common areas, a combination of Hb and DNA testing is best. However, healthcare professionals must diagnose and treat thalassemia, as proper management relies on accurately identifying the underlying condition.

A clinical update of compound heterozygosity for hemoglobin Hekinan II [a27(B8)Glu-Asp; HBA1: c.84G>T] variant in China.

Hemoglobin (Hb) Hekinan II (A27; Glu-Asp) is an α-chain variant, and its interaction with the common Southeast Asian (--SEA/) α-thalassemia (α-thal) deletion is rarely reported. This study provides a clinical update of Hb Hekinan II associated with (--SEA/) α-thal. A total of 11 simple heterozygotes and 20 composite heterozygotes for Hb Hekinan II and (--SEA/) α-thal were included based on molecular diagnosis. Hb Hekinan II exhibited a significant increase in hemoglobin, mean corpuscular volume, and mean corpuscular hemoglobin content, but a decrease in red blood cell level compared with α+ thalassemia deletion. Compared with (--SEA/) α-thal, composite heterozygotes for Hb Hekinan II and (--SEA/) α-thal showed similar erythrocyte parameters. Both heterozygotes with and without (--SEA/) α-thal showed low Hb A2 level. Hb Hekinan II showed abnormal performance in high-performance liquid chromatography but not in capillary electrophoresis. Hb Hekinan II is a benign Hb variant. The heterozygotes exhibit clinically asymptomatic coinheritance with (--SEA/) α-thal having comparable hematological phenotype to simple (--SEA/) α-thal. The combination of hematological and molecular analysis helped to improve the detection rate of this rare variant.

An Immunological and Molecular Study to Investigate the Genes (β-globin and HBA1F) in Patients with Thalassemia in Najaf Governorate

Abstract BACKGROUND: Thalassemia, a hereditary blood disorder, poses a significant health challenge globally. This study delves into the prevalence and characteristics of thalassemia, emphasizing the need for enhanced awareness and diagnostic measures, particularly in regions with high prevalence. The primary objective is to comprehensively investigate thalassemia, focusing on genetic mutations and hemoglobin (Hb) dynamics. The specific goals include identifying gene mutations through conventional polymerase chain reaction (PCR) and exploring the correlations between different Hb types. METHODS: Blood samples, totaling three milliliters for ethylenediaminetetraacetic acid tubes and 2 ml for gel tubes, were collected from thalassemia patients and a control group. Genomic DNA was extracted for conventional PCR analysis targeting α and β-globin (HBB) genes. The specific primers were designed, and PCR products were electrophoresed for the gene detection. Demographic characteristics were recorded, and Hb electrophoresis was performed to assess HbA, HbA2, and HbF levels. RESULTS: The study revealed the presence of the HBB gene in 100% of thalassemia patients causing beta-thalassemia, contrasting with the control group. The absence of the HBA1-F gene in alpha-thalassemia patients was observed. Demographic analysis showed no significant age or gender differences between the two groups. Thalassemia patients, analyzed alongside the control group, exhibit elevated HbA2 levels (2.7%–6.6%) and increased HbF levels (2%–12.1%), surpassing normal ranges (HbA2: 2%–3%, HbF: 0.8%–2%). CONCLUSION: The study confirms HBB gene association and proposes HBA1-F absence as an alpha-thalassemia diagnostic marker. Distinctive Hb patterns in thalassemia patients, highlighted by robust negative correlations (HbA-HbA2) and moderate correlations (HbA-HbF), advance diagnostic and management strategies for beta-thalassemia.

Hemoglobin Variants in Patients Attending Aden Diagnostic Center by High Performance Liquid Chromatography

Introduction: High performance liquid chromatography (HPLC) is emerging as the method of choice for initial screening and diagnosis of hemoglobinopathies. The use of alkaline and acid gel electrophoresis may result in incorrect diagnosis of hemoglobinopathies. The aim of the study is to investigate the hemoglobin pattern using the HPLC and to correlate the hematological profile with the types of hemoglobin. Methods: A cross-sectional study was conducted in Aden Diagnostic Center from July– December 2022. Over a six-month period, 250 samples of patients aged between six months to thirty years, were evaluated by HPLC for detection of hemoglobinopathies using the Lifotronic Hemoglobin Analyzer H9: β-thalassemia Analysis Mode. Red blood cell and red cell indices were determined using automated hematology analyzer. Results: A total of 152 samples (60.8%) showed different abnormal hemoglobin variants. Sixty-four (25.6%) were diagnosed to have sickle cell anemia, 46 (18.4%) as sickle cell trait, two (0.8) as beta – heterozygous thalassemia based on high level of HbA2 (&gt;3.9%), four (1.6%) as beta – homozygous thalassemia (HbF 25–91%), 18 (7.2%) as compound heterozygous state of sickle - β+ thalassemia, 17 (6.8%) as beta thalassemia trait, and one (0.4%) sample had HbD variant on HPLC was diagnosed as HbD trait. Conclusion: H9-HPLC is suitable for the routine investigation of hemoglobinopatheis because it is very powerful tool in the evaluation of Hb variants, rapid assay time and accurate quantification.

Abnormal hemoglobin anti-Lepore Hong Kong compound with β0-thalassemia ameliorate thalassemia severity when co-inherited with α-thalassemia

Abnormal hemoglobin anti-Lepore Hong Kong is a rare βδ fusion variants resulting from non-homologous crossover during meiosis. Anti-Lepore Hong Kong is known to consistently exhibit significantly increased level of HbA2. In this study, we used multiplex ligation-dependent probe amplification (MLPA) and single molecular real-time (SMRT) sequencing, as well as Sanger sequencing, to identify variants in five unrelated families with abnormal elevated HbA2 level. All probands in these five families were found to be heterozygous for anti-Lepore Hong Kong. Among them, two families showed co-occurrence of β0-thalassemia and α-thalassemia (–SEA/ or αCSα/). Heterozygotes for anti-Lepore Hong Kong displayed an average HbA2 level of 17.7% and behaved normal. However, when combined with β0-thalassemia and α-thalassemia, the probands exhibited higher HbA2 level (30.2–40.8%) and behaved with β-thalassemia trait. Furthermore, determination of the α/β-mRNA ratio revealed a slight downregulation of β-globin, similar to that of β-thalassemia minor. Our study is the first to identify compound heterozygotes for anti-Lepore Hong Kong, β0-thalassemia and α-thalassemia, provide valuable information for prenatal counseling.

SUPT5H mutations associated with elevation of Hb A2 level: Identification of two novel variants and literature review

β-thalassemia is one of the most common monogenic disorders in areas of the tropics and subtropics, which represents a major familial and social burden to local people. The elevated Hb A2 level, generally specified as greater than 3.5 %, is commonly used as a high efficiency index for screening of β-thalassemia carriers. However, mutations in other genes such as GATA1 and KLF1, could also result in increased Hb A2 level. In this study, we identified two novel variants in the SUPT5H gene: a frameshift mutation (SUPT5H: c.3032_3033delTG, p.M1011Mfs*9) and a nonsense mutation (SUPT5H: c.397C > T, p.Arg133*) in two Chinese individuals. Utilizing a combination of phenotype analysis, bioinformatics analysis, and functional analysis, we deduced that these two variants modified the SUPT5H protein's structure, thereby impacting its function and consequently leading to the heightened Hb A2 level phenotype found in the carriers. Furthermore, through a comprehensive literature review, a mutation spectrum was consolidated for SUPT5H, an investigation into the genotype-phenotype correlation was conducted, and factors known to influence Hb A2 levels were identified. Based on this in-depth understanding, clinicians are better equipped to carry out large scale screenings in regions with high prevalence of β-thalassemia.

Elevated foetal haemoglobin in tribal sickle cell anaemic patients: Blessing in disgust?

Sickle cell anaemia is a molecular disease. WHO recognises it as a global public health problem. In India, it is common among tribal communities. Increased HbS is a culprit. Therapeutic research is focused on maintaining high levels of HbF and decreasing 2,3, BPG to target disease. To assess the role of naturally compensated haemoglobin variants in tribal Sickle cell anaemic patients of North East Gujarat. Prospective, analytical, case control study conducted on randomly selected fifty tribal Sickle cell anaemic patients having disease for more than 5 years. Fifty age and sex matched, healthy control subjects. Each fifty Tribal sickle cell anaemic patients and healthy control were included in the study. Total Haemoglobin level, Sickling test by NESTROFT method and Haemoglobin variants were analysed by alkaline haemoglobin electrophoresis. Frequencies of clinical crises were recorded by oral questioning. The results were analysed using SPSS version 20. Student unpaired t- test was employed to assess the significance of the differences. P-values &amp;#60; 0.05 considered statistically significant. We observed decreased levels of total haemoglobin, high levels of HbF and HbA2 along with reduced HbA0. Compensatory increase of HbF in tribal SCA patients have shown lower frequency of clinical crises. Reduced HbA0 in sickle cell anaemia is compensated by naturally elevated HbF and HbA2 in tribal patients which has a beneficial influence on their general health.