Effect of megaloblastic anemia on hemoglobin A2 and diagnosis of ?-thalassemia trait

Context: ?-thalassemia trait is usually diagnosed by raised hemoglobin A2 (HbA2). The presence of megaloblastic anemia can cause an increase in HbA2 and create a diagnostic dilemma. Here, we have analyzed the effect of vitamin B12 and folic acid supplementation on HbA2 and diagnosis of ?-thalassemia trait in cases of megaloblastic anemia with raised HbA2. Materials and Methods: Cases of megaloblastic anemia with raised HbA2 on high-performance liquid chromatography (HPLC) were supplemented with vitamin B12 and folic acid. Post-treatment evaluation was done after 2 months. Cases showing adequate hematological response were subjected to statistical analysis. Based on post-treatment HbA2 value, the cases were diagnosed as normal, borderline raised HbA2, or ?-thalassemia trait. Pre- and post-treatment values of red cell parameters and HbA2 were analyzed. Results: There was a significant decrease in HbA2 value after vitamin B12 and folic acid supplementation. The diagnosis was changed in 70.97% of the cases after treatment. The chance of inconclusive diagnosis was decreased from more than 50% to less than 10%. Pre-treatment mean corpuscular volume (MCV) and HbA2% showed a significant difference between the thalassemic and normal groups. Conclusions: Megaloblastic anemia can lead to false-positive diagnosis of ?-thalassemia trait on HPLC. Repeat HPLC should be done after adequate supplementation of vitamin B12 and folic acid in cases of megaloblastic anemia with raised HbA2. Red cell parameters are not helpful to suspect ?-thalassemia trait in presence of megaloblastic anemia. However, HbA2% on HPLC can be a useful parameter to suspect or exclude ?-thalassemia trait in cases of megaloblastic anemia.

Anaemia in pregnancy: prevalence and treatment response to various modalities: a prospective study

Background: Anaemia seen in pregnancy are largely preventable and easily treatable if detected in time, despite this, anaemia still continues to be a common cause of maternal and perinatal morbidity and mortality in India.Methods: A prospective observational study of 200 pregnant women with anaemia was carried out from Jun 2017 to December 2018 at a Tertiary care hospital with pan India population. Patients underwent clinical examination and laboratory tests to find out the severity and type of anaemia and were treated accordingly. Iron deficiency anaemia was treated with oral or intravenous iron therapy depending upon the hemoglobin concentration. Patients were followed up after 28 days of treatment and hemoglobin estimation was done to monitor the treatment response.Results: A total 36.49% pregnant women had hemoglobin less than 10 gm%. 151 out of 200 women had serum ferritin <12 ng/ml which indicates that iron deficiency anaemia is the commonest type of anaemia in pregnancy. Overall, out of 200 patients 5.5% patients were found to have hemoglobinopathies (β thalassemia trait). After 28 days of treatment mean increase in hemoglobin was 2.40 gm% and 4.24 gm% in patients receiving oral and intravenous iron therapy respectively.Conclusions: A total 36.49% pregnant women were found to have anaemia during pregnancy and iron deficiency anaemia is the commonest type of anaemia. Therefore, there is still a need for dietary counselling and health education in the community. 5.5% patients were found to have beta thalassemia trait which was detected only after conducting hemoglobin electrophoresis. Both oral and intravenous iron therapy are effective in treatment of iron deficiency anaemia but intravenous iron therapy results in a more rapid resolution of anaemia.

Sehgal index and its comparison with Mentzer's index and Green and King index in assessment of peripheral blood smear with marked anisopoikilocytosis

Background: Mild microcytic hypochromic anaemias due to iron deficiency (IDA) and beta thalassemia trait(β-TT) continue to be a cause of significant burden to the society, particularly in the poorer developing countries. The objective of the present study was to study the RBC based indices in patients of marked anisopoikilocytosis in determining the etiology of it, to standardize few automated red cell parameters, and also objective grading of RBC morphology on peripheral smear and interpreting its utility in indicating a diagnosis. Also, to establish a relation between value of RBC indices with that of degree of anisocytosis.Methods: A total of 500 patients diagnosed with mild microcytic hypochromic anaemia on complete blood count and peripheral blood film were included in the study. Hb, RBC count, MCV, MCH and RDW obtained from the electronic cell counter were used to calculate discrimination indices by various mathematical formulae. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and Youden’s index (YI) were calculated.Results: Green and King index demonstrated the lowest sensitivity of 70.51%. Mentzer index demonstrated the highest specificity of 96.80%. The highest and lowest PPV were found for Mentzer index (97.09%) and Sehgal index (92.81%) respectively. Sehgal’s index demonstrated the highest NPV of 95.96% and lowest NPV was exhibited by G and K (87.9%). The highest and the lowest values for Youden’s index were shown by Sehgal’s index (87.82%) and G and K index (68.47%).Conclusions: Sehgal’s index followed by Mentzer index are highly sensitive and reasonably specific in differentiating β-TT from IDA and none of the indices is 100% sensitive and specific. Though HbA2 estimation is the gold standard for diagnosing β-TT, in developing countries, Sehgal index followed by Mentzer et al, index can be used to screen mild microcytic hypochromic anaemia cases to eliminate as many false positive cases as possible to reduce the financial cost.

Experience with NESTROFT for screening for thalassemia trait/ minor: evaluation against CBC and HPLC in a high prevalence region in Saurashtra, Gujarat, India

Background: Hemoglobinopathies pose a significant health burden in India. Prevention programmes can significantly reduce this burden. Although sophisticated methods of screening for β thalassemia trait are available, a cheap and simple method is beneficial for population screening. Although the Naked Eye Single Tube Red Cell Osmotic Fragility Test (NESTROFT) has been evaluated in many studies, sample sizes were small in some and many earlier studies have not done complete blood count (CBC) and High-Performance Liquid Chromatography (HPLC) in all the cases. We evaluate the suitability of NESTROFT for detection of β-thalassemia trait in a high prevalence region in Saurashtra, Gujarat.Methods: Here, 1000 unrelated individuals were studied. NESTROFT, CBC and estimation of HbA2 and HbF or other hemoglobin variants were done by HPLC.Results: Prevalence of β thalassemia trait was 7.8% in this population. NESTROFT showed an overall sensitivity and specificity of 94.87 and 85.38 respectively for the detection of β thalassaemia trait. Using red cell indices (MCH <27 pg and MCV <80 fl), One β thalassemia trait with normal indices would have been missed. Among twelve individuals with other hemoglobinopathies (HbS, HbD, HbE, δβ thalassemia trait or HPFH), seven had a positive NESTROFT while three had normal MCV & MCH values.Conclusions: NESTROFT is a cost-effective sensitive test which does not require any equipment and can be done in remote areas. It remains a useful first line screening test when large populations have to be screened.

A case of alpha thalassemia trait-associated pyoderma gangrenosum in a three-year-old Filipino male

@#INTRODUCTION: Pyoderma gangrenosum is a rare neutrophilic dermatoses in children of unknown etiology. Its occurrence may be associated with a systemic condition. In most cases, diagnosis is late and treatment is sometimes refractory to conventional therapies especially if the underlying disorder is undetected. CASE REPORT: A 3-year-old Filipino male presented with a one-year history of extensive ulcers over the nape, chest, back and lower extremities which started as a solitary painful pustule over the right gluteal area. Central healing was observed with cribriform scarring. Patient was previously treated as a case of skin infection without success. Significant laboratory findings showed severe anemia, neutrophilia, increased ferritin and increased ESR. Initial hemoglobin electrophoresis showed normal profile with mild microcytic anemia. Bacterial culture and ANA (anti-nuclear antigen) were negative. A course of oral prednisone (1-2mkd) and dapsone (2mkd) were given for a month with no improvement of the lesions. Folic acid alone was started at 5mg per day for anemia. After 2 weeks, lesions significantly improved. Three years later, upon repeat hemoglobin electrophoresis test, patient was diagnosed with alpha-thalassemia trait. Folic acid was given as mainstay therapy. Lesions healed with characteristic cribriform scarring. CONCLUSION: This is a rare case of a Filipino child who initially manifested with pyoderma gangrenosum and eventually diagnosed with alpha-thalassemia trait. Due to serious cosmetic sequelae of pyoderma gangrenosum, it is important to find and treat the underlying systemic disorder to stop progression of this debilitating and disfiguring dermatosis

Gilbert syndrome in a young boy with thalassemia trait: a rare association

Gilbert syndrome (GS) is a mild benign disease characterized by asymptomatic unconjugated hyperbilirubinemia in absence of liver disease or hemolysis. This is the most common disorder associated with bilirubin metabolism with autosomal recessive inheritance. It usually precipitates during episodes of dehydration, fasting or stress like intercurrent illnesses. Here, we are reporting a case of Gilbert syndrome in 12 yrs old boy with thalassemia trait who presented with history of persistent jaundice for last 10 months. He had disproportionately higher concentration of unconjugated bilirubin which cannot be attributed to either disorder alone. Authors considered the possibility of Gilbert syndrome after ruling out hemolytic anemia. Though genetic testing is considered to be gold standard for diagnosis of Gilbert syndrome but availability is an issue. Calorie restriction test and nicotinic acid provocation test has been used to confirm GS too. Rifampicin test, another simple test which has been described in literature though not widely used in diagnosis. It has high sensitivity and specificity too. Authors had performed rifampicin test in our index case to confirm the diagnosis of GS. Here, authors wish to highlight the patients with both GS and thalassemia trait has higher bilirubin concentrations and is more likely to be icteric than either defect alone.

Evaluation of Red Cell Indices and Discriminant Functions in the Detection of Beta Thalassemia Trait

Background: The heterozygous state of Beta-Thalassemia i.e. Beta-Thalassemia Trait (BTT) is usually not associated with any clinical symptoms and possesses abnormality in only a single BetaGlobin gene. Materials and methods: The present study was undertaken from October 2015 to October 2017 in the Department of Pathology of Mahatma Gandhi Medical College and Hospital. In our study, a total of 100 subjects were included with mild or no anemia (Hemoglobin >8 gm/dl) who were referred to the central lab for hemoglobin screening. Hb A2 values were determined by Capillary Hemoglobin electrophoresis for all 48 (BTT) and 52 (Normal/non BTT) subjects. The complete hemogram was done by automated hematology analyzer. The values of red cell indices TRBC, MCV and MCH and six discriminative indices were evaluated for detection of β-thalassemia trait. The results were compiled in tabular form and bar diagram. Results: In the present study, the patients in the BTT group had statistically significantly decreased mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) and increased TRBC count as compared to those in non BTT group. Among the six discrimination indices used highest sensitivity was found for Shine and Lal index (87.5%) followed by Mentzer index (79.16%). The highest specificity was found for E and F (88.46%) followed by RDWI (84.61%). Youden’s index was highest for Mentzer index (61.85%) followed by RDWI (55.44%). Conclusion: From this study, we thus conclude that although no screening test can diagnose β− Thalassemia Trait with 100% sensitivity or specificity, among the hematological parameters MCV, TRBC and MCH most efficiently discriminates β−Thalassemia Trait from other microcytic, hypochromic anemia. Mentzer index with CBC may be the simple, low cost, rapid and can be reliably Prachi Gupta, BP Nag, Abha Mathur. Evaluation of Red Cell Indices and Discriminant Functions in the Detection of Beta Thalassemia Trait. IAIM, 2019; 6(3): 50-59. Page 51 used as a screening test for thalassemia as a routine. However none of the formulas are 100% sensitive and specific.

Foxo3 gene expression and oxidative status in beta-thalassemia minor subjects

ABSTRACT Background: Oxidative stress may aggravate symptoms of hemolytic anemias such as beta-thalassemia. FoxO3 activation results in resistance to oxidative stress in fibroblasts and neuronal cell cultures. Objective: The purpose of this research was to study FoxO3 gene expression and oxidative status in beta-thalassemia minor individuals. Methods: Sixty-three subjects (42 apparently healthy individuals and 21 with beta-thalassemia minor) were analyzed at the Universidad Nacional de Tucumán, Argentina, between September 2013 and June 2014. A complete blood count, hemoglobin electrophoresis in alkaline pH and hemoglobin A2 levels were quantified. Moreover, thiobarbituric acid reactive species, erythrocyte catalase activity and iron status were evaluated. Beta-thalassemia mutations were determined by real-time polymerase chain reaction. FoxO3 gene expression was investigated by real-time reverse transcription-polymerase chain reaction using mononuclear cells from peripheral blood. Results: Subjects were grouped as children (≤12 years), and adult women and men. The analysis of erythrocyte catalase activity/hemoglobin ratio revealed a significant difference (p-value <0.05) between healthy and beta-thalassemia minor adults, but no significant difference was observed in the thiobarbituric acid reactive species levels and FoxO3 gene expression (p-value >0.05). Thiobarbituric acid reactive species and the erythrocyte catalase activity/hemoglobin ratio were not significantly different on comparing the type of beta-thalassemia mutation (β0 or β+) present in carriers. Conclusions: The lack of systemic oxidative imbalance demonstrated by thiobarbituric acid reactive species is correlated to the observation of normal FoxO3 gene expression in mononuclear cells of peripheral blood. However, an imbalanced antioxidant state was shown by the erythrocyte catalase activity/hemoglobin ratio in beta-thalassemia minor carriers. It would be necessary to study FoxO3 gene expression in reticulocytes to elucidate the role of FoxO3 in this pathology.

A new index to discriminate between iron deficiency anemia and thalassemia trait

BACKGROUND: The most common microcytic and hypochromic anemias are iron deficiency anemia and thalassemia trait. Several indices to discriminate iron deficiency anemia from thalassemia trait have been proposed as simple diagnostic tools. However, some of the best discriminative indices use parameters in the formulas that are only measured in modern counters and are not always available in small laboratories. The development of an index with good diagnostic accuracy based only on parameters derived from the blood cell count obtained using simple counters would be useful in the clinical routine. Thus, the aim of this study was to develop and validate a discriminative index to differentiate iron deficiency anemia from thalassemia trait. METHODS: To develop and to validate the new formula, blood count data from 106 (thalassemia trait: 23 and iron deficiency: 83) and 185 patients (thalassemia trait: 30 and iron deficiency: 155) were used, respectively. Iron deficiency, ß-thalassemia trait and a-thalassemia trait were confirmed by gold standard tests (low serum ferritin for iron deficiency anemia, HbA2 > 3.5% for ß-thalassemia trait and using molecular biology for the a-thalassemia trait). RESULTS: The sensitivity, specificity, efficiency, Youden's Index, area under receiver operating characteristic curve and Kappa coefficient of the new formula, called the Matos & Carvalho Index were 99.3%, 76.7%, 95.7%, 76.0, 0.95 and 0.83, respectively. CONCLUSION: The performance of this index was excellent with the advantage of being solely dependent on the mean corpuscular hemoglobin concentration and red blood cell count obtained from simple automatic counters and thus may be of great value in underdeveloped and developing countries.

Sehgal index: A new index and its comparison with other complete blood count-based indices for screening of beta thalassemia trait in a tertiary care hospital.

Introduction: Beta thalassemia trait (BTT) must be differentiated from iron defi ciency anemia to avoid unnecessary iron therapy and for the prevention of thalassemia major by genetic counseling. In a tertiary care hospital, it is vital that the screening tool is not only sensitive but also specifi c so as to be cost effective and save time. Aim: The aim of this study was to evaluate the new Sehgal index and compare it to existing complete blood count-based indices for the best combination of sensitivity and specifi city to predict BTT. Materials and Methods: Study was done in 2 phases - Phase 1: A retrospective analysis of 1022 consecutive high-performance liquid chromatography (HPLC) cases from July 2008 to June 2011. Phase 2: A prospective analysis of 973 consecutive HPLC cases from July 1, 2011 to June 10, 2013 was done to confi rm the results of Phase 1 and the applicability of the new Sehgal index. Results: Prevalence of BTT was 28.8% (294/1022) and 25.39% (247/973) in Phase 1 and Phase 2, respectively. Receiver operating characteristic-area under the curve and Youden index was highest for new Sehgal index, followed by Mentzers index <14. The prospective study shows results similar to those in Phase 1 confi rming the superiority of the above two indices. Conclusion: Sehgal index and Mentzers index <14 showed the best combination of sensitivity and specifi city in predicting BTT. The best indices or combination can be used as a “validated fl ag rule” in the analyzer middleware program in a hospital for identifying suspected cases of BTT.

RDW as differential parameter between microcytic anemias in "pure" and concomitant forms / RDW como parâmetro diferencial entre anemias microcíticas nas formas "pura" e em concomitância

Introduction: Iron deficiency anemia and thalassemia minor are microcytic and hypochromic types of anemia commonly found in our environment. The correct differentiation between them is of great clinical importance, although it is often hampered by the coexistence of diseases that may alter the results of standard tests for their discrimination, in addition to the significant costs of such tests. Objective: The objective of this study was to investigate the discriminatory power of red cell distribution width (RDW) between iron deficiency anemia and thalassemia minor. Method: Blood parameters were compared in 227 patients with iron deficiency anemia and/or thalassemia minor after diagnosis confirmed by molecular biology and HbA2 measurement for alpha thalassemia and beta thalassemia trait respectively. The frequency of alpha thalassemia trait in a population from two public hospitals of Minas Gerais was also determined. Result and conclusion: RDW was able to differentiate iron deficiency anemia from thalassemia trait, what indicates that this blood count parameter is a useful tool since concomitant disorders are excluded. In addition, a high frequency of the -α3.7 mutation was observed in the study population (20.3%), justifying its investigation when another cause for microcytic anemia is absent. .

Introdução: A anemia ferropriva e as talassemias menores são anemias microcíticas e hipocrômicas comumente encontradas em nosso meio. A correta diferenciação entre essas anemias é de grande importância clínica, embora muitas vezes seja dificultada pela concomitância de doenças que podem alterar os resultados dos exames padrão, além dos custos significativos de tais testes. Objetivo: O objetivo deste estudo foi investigar o poder discriminatório do índice de anisocitose eritrocitária (RDW) entre anemia ferropriva e talassemias menores. Método: Foram comparados os parâmetros hematológicos de 227 pacientes portadores de anemia ferropriva e/ou talassemia menor após diagnóstico firmado por biologia molecular ou por dosagem de hemoglobina alfa 2 (HbA2), no caso de traço alfa ou betatalassêmico, respectivamente. Foi também determinada a frequência das talassemias menores em uma amostra populacional proveniente de dois hospitais públicos de Minas Gerais. Resultado e conclusão: Observou-se que o RDW foi capaz de diferenciar as talassemias menores da anemia ferropriva, indicando que este parâmetro do hemograma é uma ferramenta útil, desde que excluídos distúrbios concomitantes. Além disso, foi verificada uma elevada frequência da mutação -α3.7 na população estudada (20,3%), o que justifica a sua pesquisa na ausência de outra anemia microcítica. .

Study of parents of β-thalassemia major children to determine cutoff values of hematological parameters for diagnosis of β-thalassemia trait and assessment of anemia in them.

Background: Because of the overlapping MCV, MCH and HbA 2 values in BTT and non-BTT subjects our laboratory determined own cutoffs. Aims: To establish cutoff values by investigating the parents of thalassemia major children and to assess the degree of anemia in BTT subjects. Materials and Methods: Study includes 179 parents of thalassemia major children (BTT cases) and 287 non-BTT controls. Samples were analyzed on an electronic hematology analyzer. The samples having MCV ≤ 76 fl and MCH ≤ 27 pg were quantified for HbA 2 by cellulose acetate electrophoresis and grey zone samples were confirmed on HPLC. Statistical Analysis Mean ± SD, sensitivity, specificity, PPV, NPV and accuracy were calculated. The histograms were plotted for MCV, MCH and HbA 2 . Results: Cases having MCV ≤ 76 fl and MCH ≤ 27 pg if considered as suspected cases of BTT then we could have missed five known BTT samples. Sensitivity increased to 100% in all three diagnostic parameters when the cutoff values were raised and specificity for MCV and MCH was decreased. But specificity was 100% with raised cutoff for HbA 2 . Hb and HCT mean values were significantly reduced in BTT cases compared to controls (P < 0.001). In 100% females and 84.9% males having BTT, mild to moderate anemia was observed. Conclusion: In our setup, the cutoff values are MCV (≤78.0 fl), MCH (≤28 pg) and HbA 2 (>3.8%) for BTT diagnosis and there is a mild to moderate anemia in BTT cases.

Hemoglobinopathies in South Gujarat population and incidence of anemia in them.

Objective: To Screen of South Gujarat population for determination of prevalence of different hemoglobinopathies particularly beta thalassemia trait (BTT) and sickle cell trait (SCT) and find out the incidence of anemia in them. Material and Methods: The present study screened 32,857 samples of students from different school and colleges in South Gujarat. Blood samples were initially tested for solubility test and complete hemogram on hematology analyzer. Samples having MCV (≤78), MCH (≤28) and/or positive solubility test were investigated for Hb electrophoresis on cellulose acetate membrane (pH 8.6). Hb A 2 level ≥3.5% was considered as diagnostic for BTT. High performance liquid chromatography on Biorad Hb variant system was done on samples having doubtful results. Result: Overall prevalence of BTT and SCT in South Gujarat was 4.4% and 1.3% respectively. Gamit, Vasava, Chaudhary, and Mahyavanshi castes had high prevalence of BTT (15.9%, 13.6%, 12.6%, and 6.9%) as well as SCT (22.2%, 15.2, 22.3, and 4.2%) respectively. Other communities like Lohana (10.8%), Sindhi (10.2%), Prajapati (6.3%), and Ghanchi (6.2%) also showed higher prevalence of BTT. Incidence of mild to moderate anemia was higher in BTT and SCT compared to non-BTT or non-SCT subjects. Conclusion : Study suggests that BTT is the most prevalent hemoglobinopathy in South Gujarat. β-thalassemia and Sickle cell anemia are highly prevalent in Mahyavanshi, Chaudhary, Gamit, Vasava and Rohit. Prajapati, Lohana, Leva Patel, and Ghanchi have β- thalassemia risk. SCT is more frequently detected in Dhodia Patel and Kukanas.

Prevalence and hematological profile of β-thalassemia and sickle cell anemia in four communities of Surat city.

Background: From the data of transfusion-dependent thalassemia major cases, the 4 communities (Muslim, Dhodia Patel, Kachhiya Patel, and Modh Bania) with high prevalence but not studied methodically were selected. Aim: The aim of this study is to find prevalence of β-thalassemia and sickle cell anemia in 4 selected communities and also to evaluate hematological profile in them. Materials and Methods: For screening of β-thalassemia trait (BTT) and sickle cell trait (SCT), all samples were tested for red cell indices, solubility, HbA 2 level and doubtful cases confirmed on HPLC. Statistical Analysis: Mean ± SD, χ2 and 't' tests were used to evaluate the significance. Results and Conclusion: Among 4 selected communities, the highest prevalence of BTT was observed in Modh Bania (6.2%) and Kachhiya Patel (6.05%) and that of SCT in Dhodia Patel (14.0%). Significantly higher prevalence of BTT was observed in Memon ( P < 0.0001) and of SCT in Khalifa 6.6% ( P < 0.0001) compared to other Muslim sub castes. Anemia was more prevalent in BTT compared to non-BTT and non-SCT subjects. 80% of Dhodia Patel non-BTT and non-SCT subjects showed microcytic red cell morphology. Their Mean ± SD Hb concentration was 12.1 ± 1.73, hence iron deficiency cannot be a sole reason. This community needs α-thalassemia and iron studies.

Perfil de beta talassemia heterozigota obtido a partir de análise data mining em banco de dados: [carta ao editor] / The profile of beta thalassemia obtained by data mining analysis in a database: [letter to editor]

Variations in the phenotypic expression of heterozygous beta thalassemia reflect the formation of different populations. To better understand the profile of heterozygous beta-thalassemia of the Brazilian population, we aimed at establishing parameters to direct the diagnosis of carriers and calculate the frequency from information stored in an electronic database. Using a Data Mining tool, we evaluated information on 10,960 blood samples deposited in a relational database. Over the years, improved diagnostic technology has facilitated the elucidation of suspected beta thalassemia heterozygote cases with an average frequency of 3.5 percent of referred cases. We also found that the Brazilian beta thalassemia trait has classic increases of Hb A2 and Hb F (60 percent), mainly caused by mutations in beta zero thalassemia, especially in the southeast of the country.

Frequency of β-thalassemia trait and other hemoglobinopathies in northern and western India.

INTRODUCTION: India is an ethnically diverse country with an approximate population of 1.2 billion. The frequency of beta-thalassemia trait (βTT) has variously been reported from <1% to 17% and an average of 3.3%. Most of these studies have been carried out on small population groups and some have been based on hospital-based patients. There is also a variation in the prevalence of hemoglobinopathies in different regions and population groups in the country. A high frequency of Hb D has been reported from the North in the Punjabi population, Hb E in the eastern region of India and Hb S is mainly reported from populations of tribal origin from different parts of the country. OBJECTIVES: To study the gene frequency of βTT and other hemoglobinopathies in three regions East (Kolkata), West (Mumbai) and North (Delhi) in larghe population group (schoolchildren) for a more accurate assessment of gene frequency for planning of control programmes for haemoglobinopathies. MATERIALS AND METHODS: This study included 5408 children from 11 schools in Delhi, 5682 from 75 schools in Mumbai and 957 schoolchildren from Kolkata who were screened for βTT and haemoglobinopathies. These included 5684 children from 75 schools in Mumbai and 5408 children from 11 schools in Delhi. Children were 11-18 years of age of both sexes. The final report is, however, only on 11090 schoolchildren from Mumbai and Delhi as data from Kolkata was restricted both in numbers and objectives and could not be included for comparison. RESULTS: The overall gene frequency of βTT in Mumbai and Delhi was 4.05% being 2.68% and 5.47% in children of the two cities respectively. In Mumbai, the gene frequency was evenly distributed. Majority of the children with βTT from Mumbai were from Marathi (38.9%) and Gujarati (25%) speaking groups. Gene frequency was >5% in Bhatias, Khatris, Lohanas and Schedule Castes. In Delhi, a higher incidence was observed in schoolchildren of North and West Delhi (5.8-9.2%). The schoolchildren of North and West Delhi comprised predominantly of Punjabi origin compared to children in the South of the city (2.2%, 2.3%). When analyzed state-wise, the highest incidence was observed in children of Punjabi origin (7.6%) and was >4% from several other states. Majority of the traits from Mumbai were anemic (95.1% male and 85.6% in female). The prevalence of anemia was lower (62.7% male and 58.4% female) children with βTT from Delhi. This was a reflection of the higher prevalence of anemia in children without hemoglobinopathy in Mumbai than in Delhi. Nutritional deficiency was probably more severe and rampant in children Mumbai. Gene frequency of Hb D was greater in schoolchildren from Delhi (1.1%) than in Mumbai (0.7%). Hb S trait (0.2%) was observed exclusively in children from Mumbai. A low incidence of Hb E trait (0.04%) was seen in children in Mumbai. A higher incidence is reported from the East. The number of cases studied from the eastern region was small as the data from the East (Kolkata) could not be included in the analysis. CONCLUSION: This study comprises a larger number of children studied for the gene frequency of βTT and other hemoglobinopathies from India. Population groups with higher gene frequencies require screening programmes and facilities for antenatal diagnosis as well as increased awareness and educational programmes to control the birth of thalassemic homozygotes. The overall carrier frequency of βTT was 4.05% and reinforces the differential frequency of β-thalassemia trait in schoolchildren from Delhi and Mumbai and the higher incidence of hemoglobin D in Punjabis as reported previously. The birth incidence calculated thereof for homozygous thalassemics would be 11,316 per year which are added each year to the existing load of homozygous thalassemics. This is much higher than the previously reported number of births annually. Hence suitable control measures need to be undertaken urgently in India.