Screening For Hemoglobinopathies Research Articles

Comparison of High-Performance Liquid Chromatography and Capillary Electrophoresis in the Screening of Haemoglobinopathies

Comparison of High-Performance Liquid Chromatography and Capillary Electrophoresis in the Screening of Haemoglobinopathies

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 (>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.

Iowa Newborn Screening Program Experience with Hemoglobinopathy Screening over the Last Two Decades and Its Increasing Global Relevance.

Hemoglobinopathies are the commonest monogenic disorder worldwide, with approximately seven percent of the world population being carriers of hemoglobinopathies. The healthcare utilization impact of thalassemia has resulted in global public health initiatives to screen for hemoglobinopathies, especially sickle cell disease (SCD). The Iowa Newborn Screening Program (INSP) has been in place for more than 50 years with a primary focus on detecting SCD. Recent changes in migration patterns have led to a global distribution of hemoglobinopathies in the western world, which has translated to an increase in the diagnosis of SCD and the incidental detection of non-sickling hemoglobinopathies within the INSP. This study documents the birth prevalence of hemoglobinopathies diagnosed in newborns through the INSP and highlights the need for newborn screening programs to evolve to meet the healthcare needs of underserved, minority populations.

Screening for Hemoglobinopathies in Pregnant Women of Haroti Region: A Guide for Surveillance, Diagnosis and Prevention

Screening for Hemoglobinopathies in Pregnant Women of Haroti Region: A Guide for Surveillance, Diagnosis and Prevention

359 Hemoglobinopathy screening in the era of expanded carrier screening: is hemoglobin electrophoresis necessary?

359 Hemoglobinopathy screening in the era of expanded carrier screening: is hemoglobin electrophoresis necessary?

Bridging the gaps in newborn screening programmes: Challenges and opportunities to detect haemoglobinopathies in Africa.

Haemoglobinopathies, including sickle cell disease and β-thalassaemia, are monogenic disorders with a relatively higher prevalence among malaria-endemic areas in Africa. Despite this prevalence, most African countries lack the necessary resources for diagnosing and managing these debilitating conditions. This study provides a critical review of newborn screening for detecting haemoglobinopathies in Africa, highlighting challenges and proposing strategies for improved diagnosis and management. A literature search on haemoglobinopathies in Africa was conducted in PubMed, Google Scholar and ScienceDirect, using specific keywords and Boolean operators, including articles published from January 1981 to December 2022. The data show that sickle cell disease is prevalent among populations in Central and West Africa; however, β-thalassaemia is prevalent among people in the northern parts of Africa. Newborn screening pilot initiatives for haemoglobinopathies were being implemented in Angola, Nigeria, Ghana, the Democratic Republic of Congo and the Republic of Benin. The cost of testing, lack of sufficient and accessible medical records, and inadequacy in healthcare infrastructure pose significant challenges in bridging the gaps in newborn screening. Furthermore, the stigmatisation and lack of awareness of haemoglobinopathies and access to newborn screening programmes pose additional challenges. This review highlights the challenges associated with haemoglobinopathy testing, effective strategies for mitigating these challenges, and future perspectives for expanding efforts toward detecting and managing these disorders across Africa. Providing affordable diagnostic tools, mobile clinics, government subsidies, education campaigns, and the implementation of electronic medical records systems could help bridge the gaps in newborn screening in Africa. The study presents a comprehensive view of newborn screening of haemoglobinopathies in Africa, provides a detailed outline of the challenges faced by newborn screening for haemoglobinopathies in Africa, and offers strategies for better diagnosis and care.

Premarital hemoglobinopathy screening program results of a province in the Black Sea region of Turkey: three years’ experience

ABSTRACT Objectives Hemoglobinopathies are a global public health problem with high mortality and morbidity and very expensive treatment. Disease can be reduced and prevented with hemoglobinopathy screening tests. It is possible to identify carriers with the hemoglobinopathy screening program applied in many countries of the world and in Turkey. This study aims to evaluate the results of the national premarital hemoglobinopathy screening program carried out in primary healthcare institutions. Methods The research is of epidemiological and cross-sectional type. Electrophoresis results examined within the scope of the premarital hemoglobinopathy screening program in Samsun between 1 January 2019 and 31 December 2021 were evaluated retrospectively. Age, gender, year of screening, and hemoglobinopathy screening results were obtained from the records. In the statistical analysis of the data, p < 0.05 was accepted. Results The median age of 52,338 people screened under the hemoglobinopathy screening program was 29.0 (16.0–86.0) years. About 54.1% (n = 28,309) of those who were screened were female, and it was found that the least screening was done in 2020 (n = 15,765 (30.1%)). As a result of the screening, the frequency of the β-thalassemia (β-thal) trait was 1.37% (n = 676), the frequency of the abnormal HbS was 0.04% (n = 20). The frequency of β-thal trait was statistically significantly higher in 2020 (1.5%) compared to other years (p = 0.029). When the results were analyzed by gender, the rate of women with abnormal HbS (3.7%) was significantly higher than the others (p = 0.017). Conclusions This study presents the results of the national hemoglobinopathy screening program in Northern Turkey and the β-thal and the abnormal HbS rates were found to be low. The data obtained will be useful in monitoring hemoglobinopathy disorders and evaluating the current program’s effectiveness in the future. It will allow decision-makers to implement policy changes and prioritize new programs.

Retrospective study on the distribution of hemoglobinopathies in Karnataka-A laboratory experience.

The importance of screening for hemoglobinopathies is well-documented in India. However, information on the distribution of hemoglobinopathies in Karnataka is lacking. The present study focuses on determining the spectrum of hemoglobinopathies for various districts of Karnataka. A retrospective analysis of samples registered for hemoglobinopathies for a period of 5 years (2017-2021) was carried out. A total of 17066 records registered only from the Karnataka region, were anonymized and retrieved. The data included gender, age, district, and results of the tests. The results were based on complete blood count, peripheral smear, and capillary electrophoresis (CE) pattern. The data were revalidated by pathologists, and the unambiguous data were analyzed for the study. One-fourth of the records (25%) showed abnormal hematological parameters. The number of female records (66%) was twice that of males and both genders showed higher distribution of thalassemia, followed by variants and double heterozygotes (DH). Several cases of thalassemia major were identified below the age of 17 years. The majority of thalassemia cases were b thal and 93% of them were b thal trait. Among the variants, HbS was more prevalent than HbE. Among the districts, Hassan had a 35.2% thal, Mysuru had a 7.2% variant, and Chitradurga had a 5.5% DH. Thalassemia, variants, and DH were distributed across several districts of Karnataka to various levels. The comprehensive retrospective analysis of the spectrum of hemoglobinopathies in various districts of Karnataka serves as evidence to carry out a prospective study on population screening where the incidence of thalassemia and structural variants is high.

Lessons learnt in the screening and diagnosis of haemoglobinopathies.

For this paper, cases reported formally and anecdotally to the authors in their screening and diagnostic roles have been selected to demonstrate areas where errors have occurred, and caution should be exercised. The cases demonstrate that it is vital that the performance and limitations of the techniques used, along with the phenotypic presentation of cases where haemoglobin variants and/or thalassaemias are coinherited are understood by those performing result interpretation. Those who deliver the service as well as those who receive reports and give results and counselling should be aware of the complexity of the topic.

Direct cord blood LAMP colorimetric phenol red assay for detecting α0-thalassemia (SEA deletion); the validation and post-natal screening in Thailand

Post-natal or newborn screening for thalassemia and hemoglobinopathies is useful for genetic counseling and managing thalassemia in children. We characterized thalassemia genotypes in newborns from the eastern part of Thailand. The results demonstrated a high heterogeneity of thalassemia and hemoglobinopathies with seventeen genotypes. We focused on α0- thalassemia (Southeast Asian [SEA] deletion) in this study. We developed and validated the loop-mediated isothermal amplification (LAMP) colorimetric assay for detecting α0- thalassemia (SEA deletion) using simple direct cord blood sampling compared to genomic DNA. A total of 160 cord blood samples were evaluated with the LAMP assay. The sensitivity and specificity of the LAMP colorimetric assay for α0-thalassemia (SEA deletion) using direct cord blood showed 100% (6/6 x 100) and 98.05% (151/154 x 100) whereas, genomic DNA showed 100% (6/6 x 100) and 100% (154/154 x 100), respectively. Moreover, we demonstrated other simple screening tools for α0-thalassemia with %Hb Bart’s, MCV, and MCH values and found that these parameters were not diagnostic in our samples. The direct cord blood with colorimetric LAMP assay is simple, rapid, and does not require a post-LAMP step compared to conventional PCR. These techniques could be applied in post-natal or large population screening for α0-thalassemia (SEA deletion). Finally, this could support early prevention of complications, early management, genetic counseling for α-thalassemia disease in children, or a long-term prevention and control program of severe thalassemia in Thailand.

Screening for haemoglobin disorders: One size may not fit all.

Accurate laboratory screening for sickle cell disease and other haemoglobin disorders is expanding worldwide. Two new reports describe different methods and strategies for screening in Mali and Denmark, respectively, and their encouraging results suggest that countries should tailor their screening programmes according to local needs, resources and opportunities. Commentary on: Guindo et al. Potential for a large-scale newborn screening strategy for sickle cell disease in Mali: a comparative diagnostic performance study of two rapid diagnostic tests (SickleScan® and HemotypeSC®) on cord blood. Br J Haematol 2024;204:337-345 and Gravholt et al. The Danish national haemoglobinopathy screening programme: report from 16 years of screening in a low-prevalence, non-endemic region. Br J Haematol 2024;204:329-336.

The Danish national haemoglobinopathy screening programme: Report from 16 years of screening in a low-prevalence, non-endemic region.

The Danish national haemoglobinopathy screening programme seeks to determine parental haemoglobinopathy carrier state antenatally. In this retrospective register-based study, we evaluated the 16-year trajectory of this programme, utilising the Danish Red Blood Cell Centre's laboratory database, covering approximately 77% of the Danish population. During the study period, we observed a substantial increase in annual diagnostic examinations performed, from 389 in 2007 to 3030 in 2022. Women constituted 88% of these cases, aligning with the emphasis of the screening programme. Of these, 54% of women of reproductive age (15-40 years) and 10% of women >40 years were specified as pregnant. During our study period, 61 children were born with a severe haemoglobinopathy, out of which 23 children were born from mothers not residing in Denmark during their first trimester thus not included in the screening programme. Prenatal invasive testing was performed for 60 fetuses, identifying 12 with homozygous or compound heterozygous haemoglobinopathy. The Danish haemoglobinopathy screening programme has provided screening, information and reproductive choices for numerous families. During the study period, screening for haemoglobinopathies has been steadily increasing and is expected to continue to increase. Awareness of and adherence to the screening programme is subject of further investigation and optimisation.

Homozygous delta beta-thalassemia: A rare case report

Delta Beta (δβ) thalassemia is a rare autosomal recessive hemoglobinopathy. This is caused by either reduced production of δ and β genes (heterozygous state) or complete absence (homozygous state). High-performance liquid chromatography (HPLC) is an important screening and diagnostic tool, to be supported by molecular data and parental studies. A 40-year-old male (propositus), with mild anemia presented in the outpatient department. There were no other definitive causes of anemia with adequate nutritional status and no response after the treatment with nutritional supplements. HPLC was advised to look for any qualitative/quantitative defects. HPLC findings reveal 96% fetal hemoglobin (HbF), a very small quantity of adult hemoglobin (HbA), and an absent HbA2. In view of this data, followed up with parental studies. Both parents had elevated HbF and similar grades of anemia. These supported the presence of a heterozygous state of δβ-thalassemia in parents and homozygous δβ thalassemia in propositus. In δβ-thalassemia, there is a deletion of the δ and β genes on Chromosome 11. Expression of the gamma gene increases to compensate, causing increased production of gamma globulin for HbF (α2 β2). On HPLC or electrophoresis, heterozygosity shows as normal HbA2 and elevated HbF, and as absent HbA2 with almost 100% HbF for homozygous individuals. Screening of hemoglobinopathies by HPLC is that can be further subjected to parental screening for a more definitive diagnosis and narrow down the differentials as compared to molecular testing considering the cost, limited availability, and higher turnaround time.

Incidental finding of rare hemoglobin: hemoglobin Bari in northeast Spain

Abstract Objectives Cation exchange high-performance liquid chromatography (HPLC) is one of the techniques available for determining glycated hemoglobin (HbA1c) and also the method of choice for structural hemoglobinopathies screening. The objective of this case is to show how in a routine HbA1c test it is possible to incidentally find a hemoglobinopathy. Case presentation In a routine blood analysis, an abnormal value for the hemoglobin A2 (HbA2) was obtained during the study of HbA1c with HPLC on the ADAMS™ A1c HA-8180T. After suspecting it could be due to the presence of a hemoglobinopathy, the study of possible variants was expanded using electrophoresis and HPLC on the Hydrasys and Variant II analysers, respectively. Since it could not be identified by these conventional methods, a genetic study was also carried out using Sanger sequencing. The patient presented a low HbA2 (1.3 %) and a 24.9 % variant with a retention time of 1.95 min, compatible with alpha-globin chain variant. In the genetic study, the pathogenic variant c.138C&gt;G was detected in the HbA 2  gene in heterozygosis, which resulted in the expression of the structural hemoglobinopathy known as hemoglobin Bari. Conclusions The initial screening for structural hemoglobinopathies allows its identification or suspicion especially when it was performed with HbA1c analysis, requiring subsequent confirmation and diagnosis by other techniques.

Thalassemia and Hemoglobinopathy Screening in Women Attending Antenatal Clinic at a Tertiary Care Center in Uttarakhand, India: A Re-look at the Laboratory Parameters Mandating High-Performance Liquid Chromatography Workup.

Thalassemia and hemoglobinopathies are the most common inherited hematological disorders. Of these, β thalassemia is the commonest disorder reported in India, followed by certain hemoglobinopathies encountered in different regions of the country. The data pertaining to the incidence of these disorders in the Uttarakhand region of India are sparse. To ascertain the prevalence and spectrum of thalassemia/hemoglobinopathies amongst antenatal women in Uttarakhand. The study also aimed to analyze the ability of red cell indices in differentiating beta thalassemia trait (BTT) from mild iron deficiency anemia (IDA). A total of 460 pregnant women in the first trimester of pregnancy were screened by cation exchange high-performance liquid chromatography. Retention time and proportions of normal/abnormal hemoglobin peaks were documented in all cases. Hemoglobin A2 (HbA2) values of ≥4% were taken as a cut-off for diagnosing BTT. Blood samples were also collected for complete blood counts, reticulocyte counts, and serum ferritin. The ability of the various discriminatory indices to differentiate between IDA and BTT was also assessed. The prevalence of BTT and hemoglobin D-Punjab trait amongst pregnant women was found to be 2.6% and 0.2%, respectively. RBC count, mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH) were found to be moderately strong predictors of BTT, with an area under the curve of 0.860, 0.857, and 0.842, respectively, which were comparable to the discriminatory indices found to be most useful in this study. In view of the 2.6% prevalence of BTT in antenatal women in this region of Uttarakhand, a routine screening will be helpful in detecting carriers early in the antenatal period. Careful interpretation of red cell indices is crucial to the distinction between BTT and IDA. Discriminatory indices are reasonably accurate in differentiating BTT from mild iron deficiency, but for practical purposes, MCV and MCH provide equivalent information to identify cases that require further workup.

Nanopore Third-Generation Sequencing for Comprehensive Analysis of Hemoglobinopathy Variants.

Oxford Nanopore Technology (ONT) third-generation sequencing (TGS) is a versatile genetic diagnostic platform. However, it is nonetheless challenging to prepare long-template libraries for long-read TGS, particularly the ONT method for analysis of hemoglobinopathy variants involving complex structures and occurring in GC-rich and/or homologous regions. A multiplex long PCR was designed to prepare library templates, including the whole-gene amplicons for HBA2/1, HBG2/1, HBD, and HBB, as well as the allelic amplicons for targeted deletions and special structural variations. Library construction was performed using long-PCR products, and sequencing was conducted on an Oxford Nanopore MinION instrument. Genotypes were identified based on integrative genomics viewer (IGV) plots. This novel long-read TGS method distinguished all single nucleotide variants and structural variants within HBA2/1, HBG2/1, HBD, and HBB based on the whole-gene sequence reads. Targeted deletions and special structural variations were also identified according to the specific allelic reads. The result of 158 α-/β-thalassemia samples showed 100% concordance with previously known genotypes. This ONT TGS method is high-throughput, which can be used for molecular screening and genetic diagnosis of hemoglobinopathies. The strategy of multiplex long PCR is an efficient strategy for library preparation, providing a practical reference for TGS assay development.

Comparison of HbA2 Using High Performance Liquid Chromatography Versus Haemoglobin Capillary Zone Electrophoresis.

Thalassemia is among the most common hereditary disorders in the world. Approximately 5% of the world's population are carriers of hemoglobinopathies, and 2.9% are carriers of beta thalassemia. Haemoglobin A2 (HbA2) constitutes less than 3% of the total hemoglobin (Hb) in adults, and the determination of Hb A2 levels is important to diagnose the beta thalassemia trait(BTT). In some cases, the level of HbA2 is not typically elevated, and some difficulties may arise in making the diagnosis. Cation exchange high-performance liquid chromatography (HPLC) and HbCZE (haemoglobin capillary zone electrophoresis) are considered acceptable methods to diagnose BTT, but these vary in their accuracy and cut-offs. In this study, we attempted to compare HbA2 values using two methods, HPLC and HbCZE, in 536 whole blood samples sent by physician-ordered hemoglobinopathy screening over two years. This included antenatal women, patients with anemia not responding to iron, and cases of familial screening where either a child or a sibling had been diagnosed with hemoglobinopathy or thalassemia. The performance characteristics of both machines were compared for the detection of the 5 most common hemoglobin variants: Hb A, HbF, HbS, Hb C, and HbE. On comparing the HbA2 values, the HPLC showed higher values for HbA2 as compared to HbCZE, while the HbF and HbS measurement agreement was good between both methods. Normal ranges and mean normal values of HbA2 differ between different methods and different manufacturers; hence, each institute using these machines should validate its cutoffs.

5613304 ANTENATAL SCREENING FOR HAEMOGLOBINOPATHIES AMONG THE TRIBAL POPULATION IN THE STATE OF TAMIL NADU, INDIA

Background: Hemoglobinopathies especially Sickle cell anemia and Thalassemia is more prevalent in the tribal population living in the hilly areas of Tamilnadu. As a part of the Government initiative for prevention of hemoglobinopathy, National Health Misiion, Tamilnadu State Chapter, has initiated Hemoglobinopathy screening for antenatal mothers in this target population. It was anticipated to have challenges like difiiculites in convincing the families for screening and if required aminocentesis. The study details with results of this screening program conducted over the period of 9 months is being presented. The project is ongoing till date. Aim: To prevent the prospective birth of Thalassemia and Sickle Cell Anemia children in Hemoglobinopathy prevalent tribal population Study Period: From September 2021 to June 2022 Methodology: All Antenatal Mothers attending the Primary health centres (PHC - Government Health Facilities) in 18 tribal blocks were screened for Haemoglobinopathies (Thalassemia & SCA) during the first trimester. 4 ml of blood was taken, 2 ml each in EDTA tubes. Informed consent was obtained before taking blood samples for test. Complete Blood Count was done in the Primary Health Centre. The second sample was transported under cold chain to the nearest medical college on the same day where high performance liquid chromatography (HPLC) was done. The CBC reports of all the mothers screened were shared with the Biochemistry Department where HPLC was done in the Biorad - 10 machine. If the mothers were found to have Thalassemia or Sickle Cell trait, husbands were tested for CBC & HPLC. If the couple was positive they were given genetic counselling regarding amniocentesis at the PHC with online support of specialists including Hematologist, State Blood Diseases Expert, and Genetics Consultant. If the couple give their consent, amniocentesis was done between 16 – 20 weeks at the prenatal diagnostic centre at state Capital, Chennai. Genetic testing was done for the parents and the foetus. This test was outsourced. If the foetus was found to be affected post test counselling was given regarding Medical Termination of Pregnancy (MTP) if they gave consent MTP was done. If the foetus was normal or to have trait, pregnancy was continued. Results: Total Mothers screened - 14016 Antenatal (AN) mothers Prevalence of Hemoglobinopathy Trait/ diseased - 7 % (959) Various types of Hemoglobinopathy trait/ diseased among the screened AN Mothers - Thalassemia trait (40.8%), Sickle cell trait (27.6 %), Sickle Cell Anemia (2%), Borderline thalassemia trait (29.6 %) No of husbands screened 511 out of which 13 % (73) were positive No of Amniocentesis done in 38 antenatal mothers out of which 31.58 % (12) were normal foetus, Trait Foetus 47.37 % (18), affected & MTP done 21.05 % (8) Conclusion: Antenatal Screening & Diagnosis is the best strategy for preventing Haemoglobinopathies which will be cost effective in the long term. It proactively reduces morbidity, mortality, DALY & psychological problems seen in management of affected children & their families. The direct medical cost of treating 1 Thalassemia child weighing 30 kg is around 2500 USD per year along with significant indirect expenses where as the cost of Antenatal Screening & Prenatal Diagnosis is 225 USD. The challenges met in implementing the program including difiiculty in taking the husband’s sample and counseling the couple with affected fetus are addressed. Reference 1. S Chakravorty, MC Dick. Antenatal Screening for Heamoglobinopathies; current status, barriers and ethics. Br J Haematol 2019; 187(4):431-440. doi: 10.1111/bjh.16188. Epub 2019 Sep 11.

Optimising the screening for haemoglobinopathies in pregnancy planning

Haemoglobinopathies are among the most common inherited disorders around the world. In the United States the diagnosis of haemoglobinopathy or a carrier state is made by universal newborn screening. However, many individuals of childbearing age do not know they are a haemoglobinopathy carrier. Screening for common haemoglobinopathies is generally offered as a part of pregnancy planning so that prospective parents can be counselled regarding the risk of having a child with a haemoglobinopathy. Multiple tests exist to screen patients for presence of haemoglobinopathy carrier or disease state; however, it is crucial to order and interpret the results correctly to appropriately counsel couples. In this case series, we describe clinical scenarios where prospective parents were surprised to unexpectedly have a child with sickle cell disease, a haemoglobinopathy that causes severe clinical complications. Through these cases we demonstrate that deficiencies in testing can occur at different levels which may lead to incorrect estimation of the risk of having a child affected by a haemoglobinopathy. Consultation with a haematologist, laboratory medicine specialist, or genetic counsellor should be considered to select the appropriate test and interpret its results.

EVALUATION OF MICROCYTIC HYPOCHROMIC ANEMIA BY ELECTROPHORESIS FOR HEMOGLOBINOPATHIES IN YOUNG POPULATION

Objectives: Hemoglobinopathies are serious genetic blood disorders requiring lifelong blood transfusions and treatment in its most severe and chronic form. If undiagnosed or untreated, these disorders ultimately lead to death. South Asia especially India, Pakistan, and Bangladesh has high populations of hemoglobinopathies. In India, there are an estimated 100,000 thalassemia majors patients and nearly 3.5 to 4 million carriers of this genetic disorder. Tribal populations in India have a range of 5–40% sickle cell anemia (SCA) sufferers. The aim of the study was to detect hemoglobinopathies by electrophoresis in microcytic hypochromic anemia in young population in the area covered by tertiary care center at Index Medical College and Hospital. Methods: A cross-sectional study was carried out in a hospital in rural area of Indore on young patients (10–30 years age group) who attended outpatient department from June 2018 to March 2020. Total 517 cases of moderate to severe anemia were included in the study. All cases were subjected to a series of hematological investigations, which include complete hemogram, peripheral blood smear, and some standard procedures. Sysmex XS800i fully automated Cell counter was used for complete blood count. It is the 5-part differential instrument. It reports on all the five subpopulations, namely, neutrophils, eosinophils, basophils, monocytes, and lymphocytes. A blood smear is a blood test used to look for abnormalities in blood cells. Results: In the study, out of 517 cases 34.4% cases were positive on Hb electrophoresis and 65.6% were negative. On electrophoresis, the number of sickle cell anemia cases was 34.3%, sickle cell trait – 20.8%, sickle beta-thalassemia – 29.8% and beta-thalassemia major – 29.8% was observed. In hemoglobinopathies, of the 73 females, 28.8% were having beta-thalassemia trait, 27.4% were having SCA 23.3% were having sickle beta-thalassemia, and 20.5% were having sickle cell trait. Of the 105 males, 30.5% were having beta-thalassemia trait, 9.5% were having sickle beta-thalassemia, 39.0% were having SCA, and 21.0% were having sickle cell trait. Conclusion: The ANMs, ASHA, and Anganwadis should be taught about the importance of screening of hemoglobinopathies. So that they can explain the purpose of screening to the female during pregnancy and to the families in the rural areas. At primary level, the CHCs and PHCs should be equipped with instruments for basic testing of hemoglobinopathies.