HbA2 Levels Research Articles

Novel variants in Krueppel like factor 1 that cause persistence of fetal hemoglobin in In(Lu) individuals

Beta-hemoglobinopathies become prominent after birth due to a switch from γ-globin to the mutated β-globin. Haploinsufficiency for the erythroid specific indispensable transcription factor Krueppel-like factor 1 (KLF1) is associated with high persistence of fetal hemoglobin (HPFH). The In(Lu) phenotype, characterized by low to undetectable Lutheran blood group expression is caused by mutations within KLF1 gene. Here we screened a blood donor cohort of 55 Lutheran weak or negative donors for KLF1 variants and evaluated their effect on KLF1 target gene expression. To discriminate between weak and negative Lutheran expression, a flow cytometry (FCM) assay was developed to detect Lu antigen expression. The Lu(a−b−) (negative) donor group, showing a significant decreased CD44 (Indian blood group) expression, also showed increased HbF and HbA2 levels, with one individual expressing HbF as high as 5%. KLF1 exons and promoter sequencing revealed variants in 80% of the Lutheran negative donors. Thirteen different variants plus one high frequency SNP (c.304 T > C) were identified of which 6 were novel. In primary erythroblasts, knockdown of endogenous KLF1 resulted in decreased CD44, Lu and increased HbF expression, while KLF1 over-expressing cells were comparable to wild type (WT). In line with the pleiotropic effects of KLF1 during erythropoiesis, distinct KLF1 mutants expressed in erythroblasts display different abilities to rescue CD44 and Lu expression and/or to affect fetal (HbF) or adult (HbA) hemoglobin expression. With this study we identified novel KLF1 variants to be include into blood group typing analysis. In addition, we provide further insights into the regulation of genes by KLF1.

“PREVALENCE OF BETA-THALASSEMIA TRAIT IN PREGNANT WOMEN ATTENDING ANTENATAL CLINIC”

INTRODUCTION- Iron deficiency anemia and thalassemia syndromes,especially beta thalassemia trait (BTT),are the two most commonly ccuring microcytic hypochromic anemias highly prevalent in countries like India. Iron deficiency anemia is the first most common cause of anemia in pregnancy,beta thalassemia trait is the second most frequent cause of anemia of pregnancy. Patients with thalassemia trait shows an increased incidence of anemia during the second trimester of pregnancy. Beta Thalassemia Trait (BTT) patients are usually asymptomatic and ignorant of their carrier state unless diagnosed by testing. Screening for thalassemia can be done by measuring HbA2 levels MATERIAL AND METHOD- This is a retrospective study. The data were collected from a clinic and associated st pathology in Shivpuri and a private maternity hospital in Gwalior from 1 January 2019 to 30 June 2021.94 patients were included in the study.All pregnant women between the age of 18-45 years and at any gestational age with hemoglobin level <10g/dL and microcytic hypochromic anemia (report of CBC) were included in the study. High-Performance Liquid Chromatography (HPLC) and serum ferritin report were collected. RESULTS- In our study the most common age group in which the anemia was found between 19-35 years (80.85%).Most of the patients were primigravida (44.68%),between 26-37 wks of gestational age (82.97%),from a rural area (70.21%). Only 2 patients had a family history (2.12%) of beta-thalassemia and 6 patients had a history (6.38%) of blood transfusion. In our study moderate anemia was found in 55.31% of cases followed by mild anemia 25.53% and severe anemia 19.14%. In our study iron deficiency anemia was found in 87.23% only minor Beta-thalassemia was found to be 4.25%, of the patient,and the coexistence of iron deficiency anemia and beta-thalassemia in pregnant women is 8.51%. In our study total of 12 patients (4 cases of only minor beta-thalassemia and 8 cases of minor beta-thalassemia associated with IDA) The prevalence rate of beta-thalassemia in our study was 7.8%. CONCLUSION- the recommendation is to diagnose beta-thalassemia in pregnant patients is essential to give proper treatment.The screening of pregnant patients prevents unnecessary parental iron therapy and iron overload in case of anemia.further studies are needed in this field and less expensive and accurate methods should come in place.

Differential Diagnosis of Three Commonest Deletion β-Thalassemia in Chinese

To analyze the hematological characteristics of Chinese Gγ+(Aγδβ)0-thalassemia，SEA-HPFH and Taiwan type β-thalassemia. Hemoglobin electrophoresis and blood routine test were used to analyze the hematological indexes of all peripheral blood samples，PCR-Flow fluorescent hybridization and Gap-PCR were used to detect the globin gene mutations and the data were analyzed statistically. The 3 types of deletion β- Thalassemia patients were showed as hypochromic small cell anemia. The MCH and MCV values of Taiwan type β-thalassemia patients were the lowest. The results of hemoglobin electrophoresis showed that the increasing of HbF was found in all of the 3 types. Except for the decreasing of Hb A2 in Chinese Gγ+(Aγδβ)0-thalassemia，the levels of Hb A2 in the other two deletion β-thalassemia patients were significantly increased. Except for Hb, there were significant differences in MCV, MCH, Hb A2 and HbF between Chinese Gγ+(Aγδβ)0-thalassemia and SEA-HPFH(P<0.001). Through analyze the hematological characteristics, it can be provide that the guidance for the differential diagnosis and genetic consultation of the three commonest deletion β-thalassemia in Chinese.

Microcytic Anemia and Celiac Disease Preceding Pulmonary Symptoms in Lane-Hamilton Syndrome

History and Case PresentationPatient is a 36 year old white male with a history of chronic microcytic anemia and tobacco use. He could not recall any pertinent family history. His chronic anemia started at age 29, at which time he had no pulmonary symptoms. Representative laboratory studies included HGB 7.7 HCT 27.2 MCV 60.7 FE/TIBC of 13/428 with albumin of 3.2. He was diagnosed with iron deficiency, and received multiple intravenous iron infusions. These therapies briefly increased his hematocrit, however, he consistently required PRBC transfusions within 1-2 months of receiving intravenous iron, as he would again become anemic with microcytosis; he estimated that he had received approximately 30 units of PRBCs since age 29. Dyspnea on exertion was first noticed at age 34, which progressively worsened and was associated with occasional episodes of blood tinged sputum and cough, but never frank hemoptysis. Pulmonary function tests in 2016 were reportedly normal, and there were no abnormalities on CXR.Multiple EGDs and colonoscopies were performed without any evidence of bleeding. Duodenal biopsies with villous flattening were first noted in 2014. Patient adhered to a gluten free diet for two months immediately following his diagnosis of celiac sprue and reported that his anemia stabilized, however he stopped the diet for unclear reasons.He presented to our facility in May of 2017 with intermittent sharp chest pain, severe dyspnea on exertion, and lightheadedness. Initial HGB was 7.2 HCT 24.7 MCV 59.2 WBC 6700 PLT 104000. Iron studies obtained prior to transfusion reveal Fe/TIBC of 17/253, ferritin 90, albumin 1.8. Peripheral smear was consistent with iron deficiency, with mild to moderate anisopoikilocytosis. Absolute reticulocyte count the day after admission was 0.104 with HCT of 21.7.He had a similar presentation 8 months prior, in October 2016. CT chest was remarkable for ground-glass opacities, pulmonary fibrosis, emphysema, and mild bilateral hilar adenopathy. There was nephrotic range proteinuria, and albumin was 1.9. He was given ferric gluconate with improvement in his symptoms, and was discharged four days later with HGB of 9.2. He was lost to follow up until May, and did not receive scheduled EBUS and lung biopsy. Repeat CT scan was performed on second admission, showing interval progression of peripheral fibrosis and honeycombing (Image 1).Investigations in 2017 included an EGD/colonoscopy, with single duodenal biopsy finding of increased intraepithelial lymphocytes, with preserved villous architecture, h. pylori negative. Tissue transglutaminase IgA was >250 U/mL and anti-gliadin IgA was 26 U/mL. Genetic testing revealed HLA DQ2 serotype. There was no evidence of CHF or pulmonary HTN on Echo. Hemoglobin electrophoresis revealed no abnormal hemoglobin migration and physiologic HbA2 and HbF levels. B12 and folate levels were within normal limits. Lung biopsy revealed alveolar filling with numerous hemosiderin laden macrophages, in background of alveolar pneumoconiosis and phlebosclerosis, (image 2, high magnification).He was administered PRBCs and ferric gluconate, and discharged with instructions to initiate gluten-free diet, as well undergo renal biopsy to evaluate nephrotic syndrome.DiscussionLane-Hamilton Syndrome (LHS), the association of Idiopathic Pulmonary Hemosiderosis (IPH), celiac disease, and microcytic anemia is an extremely rare clinical entity with 15 worldwide cases described in adults. When patient presented with microcytic anemia, ferritin was consistently in the normal range, while remaining iron studies including Fe/TIBC and transferrin saturation were consistent with iron deficiency when corrected for proteinuria and transferrin loss. Historic CBC and iron studies revealed a clear iron deficiency pattern as described. Parenteral iron infusion did improve hematocrit on multiple presentations, but response was transient. Celiac disease was proven by biopsy and serology. Hemosiderin deposition was found on lung biopsy without a plausible alternate etiology. These features are consistent with known characteristics of LHS. While iron deficiency resistant to iron supplementation has been described without inital pulmonary findings in children with IPH, to our knowledge, iron deficiency with celiac disease preceding the development of IPH by such a long interval has not heretofore been described in adult LHS. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Red cell parameters, HbA2 levels, and the best discrimination index in beta thalassaemia trait in Sri Lanka

Introduction: Beta thalassaemia trait (BTT), also known as thalassaemia carrier state, leads to hypochromic microcytic indices. Identifying asymptomatic carriers on full blood count will help medical personnel to direct them for confirmatory tests such as High Performance Liquid Chromatography (HPLC) and prevent birth of babies with thalassaemia major by discouraging intermarriage between two individuals with BTT. Objectives: To describe red cell parameters and HbA2 levels of individuals with BTT in Sri Lanka and determine the best discrimination index for screening of BTT. Methods: BTT confirmed by HPLC (HbA2 ≥3.5%) during a two year period at the Medical Research Institute, Colombo, Sri Lanka were selected and full blood count (FBC) reports were analysed. Haemoglobin (Hb), RBC parameters; RBC count, MCV, MCH, MCHC, and RDW and HBA2 of the study population was analysed and the mean and the standard deviation (SD) was calculated for each parameter. These parameters were then analysed using Mentzer, Srivastava, England and Fraser, Shine and Lal, Green and King and Thal Index and sensitivity of each index for detecting BTT was calculated to ascertain which index identified BTT with maximum sensitivity. Results: 157 BTT cases without concurrent iron deficiency and 206 BTT without serum ferritin results were analysed. In the BTT group without iron deficiency, Hb was 8.3-11.1 g/dl; mean 9.7 g/dl, RBC count 4.1-5.7; mean 4.9. MCV 59.2-68.8 fl; mean 64 fl. MCH 18.1-21.9 pg; mean 20 pg. MCHC 29.4-32.6 g/dl; mean 31 g/dl. RDW 12.2-23% mean 17.6%. HBA2 level for BTT in Sri Lanka was 4.8% (SD 0.5). Sensitivity for each index was, Mentzer Index 57.1%; England and Fraser Index 25.3%; Srivastava Index 44.2%; Green and King Index 61.5% Thal Index 73.3%; Shine and Lal Index 98.7% and New index was 92.3%. Conclusions: All RBC parameters except MCHC were outside the normal ranges. A mean MCV of 64 fl (SD 4.8) and MCH of 20 (SD 1.9) best denoted BTT on FBC. The lower limit of HbA2 (4.3%) in BTT in Sri Lanka is slightly higher than the currently used diagnostic value of ≥ 3.5%. Shine and Lal Index with a sensitivity of 98.7% was the most reliable index to identify BTT in Sri Lanka. Vigilant application of above findings on FBC by first contact doctors will help to identify more thalassaemia carriers.

A Compound Heterozygous HBB: C.79G&gt;A Associated with HBB: C.-78A&gt;G Mutation of HBB Gene Leading to Β-Thalassemia Intermedia in A Vietnamese Patient: A Case Study

β-thalassemia is an inherited blood disorder that is the most common single-gene disorder worldwide and in Vietnam. This study reports an intermedia β thalassemia patient in a family from Northern Vietnam. The mother has compound heterozygous thalassemia presenting mutation of hemoglobin HBB: c.79G>A associated with HBB: c.-78A>G. The result indicates that the determination of molecular characterization of subjects presenting normal HbA2 level associated with abnormal red cell indices is needed for accurate diagnosis and improved genetic counseling.

HbA2 induction: the merit of pancellularity in sickle cell disease.

HbA2 induction: the merit of pancellularity in sickle cell disease.

Elevated Hb A2 is Not Always Indicative of β-Thalassemia

Hb A2 levels are usually high in carriers of β-thalassemia (β-thal). These levels also provide a sensitive marker for the identification of hemoglobin (Hb) variants. In this study, we aimed to examine two patients from two Chinese families who showed elevated Hb A2 levels but did not show any signs of β-thal. The HBB variants were analyzed using direct sequencing of HBB and in silico prediction analysis. Moreover, the family’s genetic history was investigated. We examined two probands from different Chinese families with elevated Hb A2 levels who were not afflicted with β-thal, although several nucleotide changes were found at codon 81 (CTC>CTA) (HBB: c.246C>A) in Family 1 and a compound heterozygosity for codon 40 (AGG>AAG) (HBB: c.122G>A) and IVS-II-478 (C>A) (HBB: c.316-373C>A) in Family 2. After investigating the genetic history of both families including the β-thal aspect, we found that these mutations were not responsible for the elevated Hb A2 levels. It is rarely reported that high Hb A2 level is not indicative of β-thal. In contrast, low or normal Hb A2 level is always found with β-thal due to other molecular defects that mask their β-thal genotype. Our results highlight the importance of considering the genetic factors related and unrelated to β-thal to improve the accuracy of future genetic counseling.

Detection of the HBB: c.393T>G Mutation in Two Patients with Hypochromic Microcytic Anemia

A novel mutation, HBB: c.393T>G on the HBB gene, was detected in two hypochromic microcytic anemia patients from Yulin, in the Guangxi Province of the People’s Republic of China (PRC), by next-generation sequencing (NGS). It is a nonsense mutation causing a stop codon at amino acid 131 in exon 3 of the HBB gene. It was found in a heterozygous state in two patients who both presented severe anemia during pregnancy and moderate anemia before pregnancy; Hb A2 levels were slightly increased (more than 4.0%) in both patients. It was also detected in the father of one of the patients. This mutation was pathogenic, and caused the dominant thalassemia-like phenotypes in the two patients.

Fetal-haemoglobin enhancing genotype at BCL11A reduces HbA2 levels in patients with sickle cell anaemia.

Understanding the interplay of genetic factors with haemoglobin expression and pathological processes in sickle cell disease is important for pharmacological and gene‐therapeutic interventions. In our nascent study cohort of Nigerian patients, we found that three major disease‐modifying factors, HbF levels, α‐thalassaemia deletion and BCL11A genotype, had expected beneficial haematological effects. A key BCL11A variant, while improving HbF levels (5.7%–9.0%), also led to a small, but significant decrease in HbA2. We conclude that in general, interventions boosting HbF are likely to reduce HbA2 in patients’ erythroid cells and that such therapeutic strategies might benefit from a parallel stimulation of HbA2 through independent mechanisms.

Changes in HbA2 Levels May Signify Hemoglobin Defects in Infants

Changes in HbA2 levels may signify hemoglobin defects in infants

Selective electrochemical sensing of hemoglobin from blood of β-thalassemia major patients by tellurium nanowires-graphene oxide modified electrode

Hemoglobin is an important part of the biological system that is involved in the transportation of oxygen and carbon dioxide. Abnormal level of hemoglobin is associated with different diseases such as β-thalassemia and anemia. An electrochemical sensor based on graphene oxide-tellurium nanowires (TeNWs/GO) is developed for the first time for electrochemical sensing of hemoglobin (Hb). Sensing parameters such as concentration, pH, scan rate, and effect of interferences are optimized. Linearity, detection, and quantification limits for TeNWs/GO-GCE are 0.996, 0.29 μM, and 0.967 μM respectively. Stability is measured by cyclic voltammetry (100 cycles) and chronoamperometry (15 h). TeNWs/GO-GCE quantitatively determines hemoglobin from blood samples of β-thalassemia major patients. To evaluate the relationship between RBC’s count and hemoglobin β-thalassemia major patients with the level of HbA, HbA2, and HbF statistical analysis is carried out. Shapiro-Wilk analysis shows a non-normal distribution and the results are calculated by Spearman correlation analysis. The results show that thalassemia patients have low levels of hemoglobin due to mutations in globin chains of hemoglobin.

In silico prediction of HBD gene variants in the Iranian population

BackgroundThe quantification of hemoglobin A2 (Hb A2; α2δ2) is used as a valuable test to differentiate α- and ß-thal carriers in clinical laboratories. Therefore, the HBD (δ-globin) gene variants could result in reduced levels of Hb A2 and have implications for thalassemia screening programs. The aim of the present study was to predict the consequences of HBD gene variants identified in the Iranome project.ResultsThe highest number of variants was in the Persian Gulf Islanders. The variants of p.Gln132Glu (HBD: c.394C>G), p.Gly17Arg (HBD: c.49G>C), p.Thr5Ile (HBD: c.14C>T), and p.Ala28Ser (HBD: c.82G>T) presented damage results in three or more prediction tools. In addition, it seems that the p.Gly30= (HBD: c.90C>T) decreases the use of authentic splice and, instead, creates a new donor splice site (DSS) or leads to the use of a cryptic DSS.ConclusionsMost of these variants have been associated with a decrease in Hb A2 levels. Due to the high mutational diversity in the HBB gene in the Iranian population and the use of Hb A2 quantification to differentiate α- and ß-thal carriers among Iranian clinical laboratories, some attention should be taken to a possible co-inheritance of HBD gene variants to avoid the misdiagnosis of ß-thal carriers.

β-globin gene cluster mutations and large deletions among anaemic patients with elevated HbF level in a tertiary teaching hospital in Kelantan, Malaysia

Mutations in the β-globin gene cluster can lead to β-thalassaemia, δβ-thalassaemia, hereditary persistence of foetal haemoglobin (HPFH) and some of the haemoglobin variants. The clinical and haematological spectrum of thalassaemia range from benign to pathogenic conditions including severe transfusion dependent thalassaemia. Therefore, it is important to accurately diagnose β-globin gene cluster mutations to prevent thalassaemia major or intermedia offspring. The objective of this study is to detect β-globin gene cluster mutations and large deletions among anaemic patients with elevated HbF level in a tertiary teaching hospital in Kelantan, Malaysia. This study involved 144 anaemic patients with HbF level &gt;1.0%. High-Performance Liquid Chromatography (HPLC) was used to determine the HbF and HbA2 levels. Multiplex-ARMS (ARMS)-PCR and gap-PCR were performed for those patients with high HbA2 level (&gt;3.2%) and normal HbA2 level (≤3.2%) to detect β-globin gene cluster mutations and large deletions respectively. The majority of patients were Malays (99.3%) with mean age of 19.99 ± 1.64 years and female 61.1% predominance. Out of 87 samples tested using multiplex ARMS-PCR against eight targeted single mutation; Cd 41/42, IVS 1–5, Cd 26, Cd 17, Cd 71/72, IVS 1–1, Cd 8/9 and -28 mutations, 65 (74.7%) patients were detected which comprises of Cd 26 (56.3%), Cd 41/42 (11.5%), compound Cd 26 and Cd 41/42 (3.4%) and IVS 1–1 (3.4%). Meanwhile, for multiplex gap-PCR which detect four types of large deletions; Thai (δβ)o-thalassaemia, HPFH-6, Siriraj J and Hb Lepore, one out of 57 patients (1.8%) was found positive with Thai (δβ)o-thalassaemia. There was a significant difference between the mean of HbF level, MCV level as well as MCH level of patients with and without β-globin gene mutations and large deletions (P&lt;0.05). This study highlighted the presence of various types of β-globin gene cluster mutations detection in establishing a definitive diagnosis among this selected group of patients for the large-scale screening of the thalassaemia gene.

Evaluation of Naked Eye Single Tube Red Cell Osmotic Fragility Test (NESTROFT) in Screening and Diagnosis of Beta Thalassaemia Trait in Pakistani Population

Background: Beta thalassaemia is an inherited disease that is a cause of continuous burden on the affected families and the society. The objective of the study was to evaluate the efficacy of Naked Eye Single Tube Red Cell Osmotic Fragility Test (NESTROFT) for the detection of beta thalassaemia trait when used for the screening in Pakistani population. Methods: All participants requesting a complete blood count were included in the study. Informed consent was obtained. Naked Eye Single Tube Red Cell Osmotic Fragility Test was performed with freshly prepared 0.36% buffered saline. Turbidity in the tubes was an indication for individuals to be positive for beta thalassaemia trait. Hemoglobin electrophoresis was performed on all positive cases to confirm the sensitivity and the reliability of the screening test. Chi-Square test was used to test for any significant correlation between the different hematological parameters and other variables. Results: A total of 812 patients were included in the study from January 2017 to June 2017. The mean age of participants was 35.5 + 13.6 years. There were 351 (43.2%) male and 461 (56.7%) female participants. A 100% Naked Eye Single Tube Red Cell Osmotic Fragility Test sensitivity was obtained with 65 (8%) positive cases having HbA2 levels of more than 3.2% indicating their true positivity whereas, all 747 (92%) negative cases having HbA2 levels of less than 3.2% indicating their true negativity. Conclusion: NESTROFT was found and a rapid screening test for the identification of carriers of beta thalassaemia trait in our population.

Evaluation of Beta Globin Gene Mutations in Beta Thalassemia Carrier Children in Aydın Province and its Environment

Objective: Beta thalassemia carriers (BTC) in Turkey is observed with a frequency of 2.1%, and it is the most common cause of anemia after iron deficiency. There are few studies showing the effect of genotype on phenotype in beta thalassemia carrying children. The aim of this study is to determine the mutation diversity of these children in and around Aydın and evaluate the effects of these mutations on complete blood count parameters and hemoglobin electrophoresis. Methods: This study included mutation analysis of 65 patients who were diagnosed as BTC in Adnan Menderes University, Faculty of Medicine, Department of Child Health and Diseases, Pediatric Hematology Outpatient Clinic between 01.01.2014 and 01.08.2019. Complete blood count, hemoglobin electrophoresis and mutation analysis results were obtained from the computer data system and patient files. Research data were evaluated by using SPSS 21.0 statistics program. Results: 39 (60.0%) of the sixty-five patients were male, 26 (40.0%) were female, and the mean age was 8.34±4.94 years. When full blood count parameters were analyzed, 87.7% had anemia, 100% microcytosis and high red cell distribution width (RDW) level, 49.2% hypochromia and 87.7% high red blood cell (RBC) count. RDW level was ≥16 in 66.2% of the cases. Seventeen different mutations were detected. The mutations most frequently occurred in “intron 1” gene region (66.1%). The most common ones were IVS I-110 (44.11%), IVS I-1 G&gt;A Heterozygotes (8.8%,) IVS I-6 T&gt;C Heterozygotes (7.5%) and IVS II-745 (7.5%). RDW level was ≥16 in 66.2% of the cases. Seventeen different mutations were detected. The mutations most frequently occurred in “intron 1” (66.1%) gene region. IVS I-110 (44.11%), IVS I-1 G&gt;A Heterozygotes (8.8%,) IVS I-6 T&gt;C Heterozygotes (7.5%) and IVS II-745 (7.5%) were observed most commonly. In patients with IVS I-110 mutation, mean Hb level was 10.55 gr/dL, MCV level was 58.44 fL, RDW level was 16.51, RBC level was 5680x10⁹/L, HbA2 level was 4.77%, HbF level was 2.34%. Mutations detected in 12 patients with HbF level above 5% were as IVS I-110 (5 people), IVS I-6 T&gt;C Heterozygotes, Codon 39 C&gt;T Heterozygotes, IVS I-116, c.25-26 del AA (plys9Valfs) Heterozygotes, c.27dupG (pSer10valfs*14), c316-373 (IVSII-478 C&gt;A Heterozygotes), -87 C&gt;T Heterozygotes. Mentzer index was calculated as &gt;13 in six patients (9.2%). The mutations seen in these patients were IVS-I 110, c.27dupG (p.Ser10valfs*14), c316-373 (IVS II-478 C&gt;A Heterozygotes), -87 C&gt;T Heterozygotes. There were four patients (6.1%) with a RDW index &gt;220. Two of these patients had c.27dupG (p.Ser10valfs*14) and others had c316-373 (IVS II-478 C&gt;A Heterozygotes) and -87 C&gt;T Heterozygotes mutations. Mutations detected in four patients with HbF levels in the range of 9.48-15.67 were IVS I-116 T&gt;G Heterozygotes, IVS I-110 G&gt;A Heterozygotes, c.25-26 del AA (p.lys59valfs) Heterozygotes, c27 dupG (pSer10 valfs*14) and three of these mutations carrying β° mutation type were located in exon 1 and one of them carrying β⁺ mutation type (IVS I-110) was in intron 1. Conclusion: The same mutations detected in patients with beta thalassemia carriers have different effects on complete blood count parameters. HbA2 and HbF levels suggest that these mutations are not effective on the phenotype alone and there may be additional factors which should be clarified. We think that there may be BTC in cases with low RBC, Mentzer index ≥13 and RDW index ≥220, HbA2 &lt;3.5 and studying the mutation analysis of these patients will contribute significantly to the literature.

Background: Iron Deficiency Anemia (IDA) and beta-thalassemia trait (&amp;beta;-TT) are the two most prevalent causes of microcytic hypochromic anemia. The current study aimed to assess the accuracy of the various indices to distinguish these two conditions. Material &amp;amp; Methods: A total of 40 patients were studied retrospectively by calculating six discrimination indices. The patients were assessed according to the Mentzer,

Background Iron Deficiency Anemia IDA and beta-thalassemia trait beta-TT are the two most prevalent causes of microcytic hypochromic anemia. The current study aimed to assess the accuracy of the various indices to distinguish these two conditions.Material amp Methods A total of 40 patients were studied retrospectively by calculating six discrimination indices. The patients were assessed according to the Mentzer Shine amp Lal England amp Fraser Green amp King Red Cell Distribution Width Index RDWI and Srivastava indices. The study included 19 beta-TT cases based on low Hb and mean cell volume MCV levels normal serum iron and raised levels of HbA2 gt3.5 and 21 IDA cases with low hemoglobin low serum iron low ferritin high total iron-binding capacity and normal electrophoresis. The accurately diagnosed cases were identified and their sensitivity specificity positive and negative predictive value and Youdenrsquos index of the indices were calculated.Results None of the indices showed a sensitivity and specificity of 100. The percentage correctly diagnosed was highest for RDWI 97.5 followed by Green and King 95. Youdenrsquos index was also highest for RDWI 95 followed by Green and King 90 and Mentzer 84.7.Conclusion RDWI Green amp King and Mentzer are readily available and the most stable indices in differentiating between IDA and beta-TT.

Description of hemoglobin H disease mutations in alpha thalassemia patients in Sulaimani Region in Kurdistan Region, Iraq

CONTEXT : Hemoglobin H (HbH) disease is induced by mutations in three out of the four α- globin genes. Most commonly, mutations are either deletional or nondeletional. While some deletions (3.7 and 4.2) induce α+ thalassemia, others induce (20.5, MED, THA1, FIL) α0 thalassemia. HbH disease is a combination of both.AIMS : This study aimed to describe alpha-thalassemia (HbH disease) mutations in Suliamaniyah Province, Iraq.MATERIALS AND METHODS : Fifty-one patients with hypochromic microcytic anemia were evaluated for HbH disease. For each patient, a 2-ml venous blood sample was taken for isolating DNA. The samples were inspected for HbH disease mutations by gel electrophoresis, applying the α-Globin Strip Assay from the Vienna Lab TM commercial kit.STATISTICAL ANALYSIS: Microsoft Excel software was used to analyze data.RESULTS : Clinical data from complete blood count, hemoglobin (Hb)-electrophoresis, and HbH test were measured. HbH patients had significantly low levels of mean corpuscular volume, mean corpuscular Hb, and Hb (HGB) compared to normal values, and all showed a positive result in the HbH test with a low level of HbA2. Both the Med double gene deletion (3.7/MED) and the 3.7 single-gene deletion were detected in 68.62% of patients. Single-gene deletion 4.2, double gene deletion 20.5 (4.2/20.5), double gene deletion Med, and point mutation α2 poly A2 (MED/α2 poly A2) were all found in 1.96% of patients.CONCLUSION : There is no difference between the phenotypes of patients with different genotypes.

Hemoglobin fractions in Indian pediatric population – Do we need to look westward?

BACKGROUND: Hemoglobin (Hb) is a tetramer of two alpha and two beta globin polypeptide chains, The fractions of HbF, HbA2, and HbA vary gradually in pediatric population (age 0–12 years) and show a dynamic change in the 1st year of life.OBJECTIVE: The objective of the study is to establish the levels of normal hemoglobin (Hb) fractions by using high-performance liquid chromatography (HPLC) in Indian pediatric population.MATERIALS AND METHODS: A total of 169 children of 0–12 years of age were recruited from a pediatric outpatient clinic. Eleven cord blood samples from normal deliveries were collected, and 2 ml peripheral blood was drawn from each subject in EDTA vial for CBC and Hb HPLC. CBC was performed by hematology analyzer XT 2000i, and the proportion of HbA, HbA2, and HbF was obtained from HPLC using the β thalassemia short program.RESULTS: The fractions of HbF, HbA2, and HbA gradually changed with increasing age. HbF levels decreased rapidly from 80.9% ± 0.48% (mean ± standard deviation) in the cord blood to 3.6% ± 1.04% at 6 months of age. HbA2 was 0% in cord blood, was 0.04 ± 0.12 in newborn, reached to a level of 2.52% ± 0.3% at 6–12 months of age, and thereafter marginally increased to 2.65 ± 0.89% at 2–12 years of age. HbA was 20.41% ± 5.14% in the cord blood and increased substantially to 84.7% ± 1.83% at 6–12 months of age.CONCLUSION: HbF levels show a more dynamic change in the first 6 months of life than later half. It reaches adult levels by the age of 2 years. HbA2 levels reach a plateau at 6 months of age. HbA levels rise substantially until the 6th month and sustain thereafter. These data can serve as a quick practical reference guide for the analysis of Hb fractions in the Indian pediatric population.

NESTROFT AS SCREENING TEST FOR BETA THALASSEMIA TRAIT

Objective: To evaluate the utility of NESTROFT as a screening test for Beta Thalassemia Trait. Material and Methods: A Prospective study of total 137 clinically normal siblings of thalassemia major patients registered at thalassemia clinic in department of pediatrics, Govt. medical college, Kota were selected randomly for the study. All cases were investigated for CBC, Mentzer’s index, NESTROFT and HbA2 estimation by HPLC method. Result: Total 137 cases were screened for thalassemia carrier by HbA2 estimation using HPLC technique. Total 67 (48.90%) cases having HbA2 level ≥3.5 % considered as carrier whereas rest 70 (51.10%) cases were normal. In this study 95 cases showed MCV&lt;80. Out of them 59 cases were carrier and 36 cases were normal. The Sensitivity, specificity, positive predictive value and negative predictive value of MCV&lt;80 are 90.12%, 49.71%, 63.78% and 84.68% respectively. The Sensitivity, specificity, positive predictive value and negative predictive value of Mentzer index are 85.25%, 97.16%, 96.45% and 89.19 % respectively. In present study, NESTROFT were done in all cases, the sensitivity of NESTROF test is 94.21%, specificity is 89.72%, Predictive value of positive test is 89.17% and Predictive value of negative test is 93.75%. So NESTROFT is better test for screening for Beta Thalassemia trait. Conclusion: NESTROF test is reliable, cost effective and better screening test for Thalassemia carrier detection. Positive cases can be confirmed by HPLC. NESTROFT can be used as mass screening programme for hemoglobinopathies.