HbE Trait Research Articles

Reproducibility of Naked Eye Single Tube Red cell Osmotic Fragility Test (NESTROFT) as a screening test for the detection of b thalassemia and Hemoglobin E traits with the variation of reading time

Background: Though 0.36% NaCl buffered solution has been used as an effective reagent for carrying out Naked Eye Single Tube Red cell Osmotic Fragility Test (NESTROFT) as a screening test for the detection of b thalassemia and Hemoglobin E traits, reading time for its interpretation is yet to be settled. Methods: This cross -sectional analytic study was done at Department of Pediatric Hematology and Oncology, BSMMU from January 2006-June 2007. NESTROFT was performed in 3 separate tubes for each of the 100 subjects (with â thalassemia trait and HbE trait and normal hemoglobin) with 0.36% NaCl buffered solution and reading was taken at different time at 5-minute, 15-minuteand 30-minute. Results were recorded as positive and negative. Hb electrophoresis, Hemogram with RBC indices for each sample and iron profile of selected subjects were done. Results: Among the 100 study subjects, 48 were carriers (18 were â thalassemia traits, 30 were Hb E traits). Age range was 1 to 52 yrs. Mean Hemoglobin concentration, MCV, MCH of both b thalassemia and Hb E carrier were significantly lower than normal subjects (p values <. 001). At 5-minute observation, NESTROFT was positive in 17 out of 18 â thalassemia traits with a sensitivity of 94.4%, 24 out of 30 HbE traits gave positive result with a sensitivity of 80%. Among the 52 normal subjects 35 gave negative result with a specificity of 67%. Positive predictive value and negative predictive value for detection of b thalassemia traits and HbE traits were 50% & 97% and 58% and 85%respectively. Result of NESTROFT interpreted at 15-minute and 30-minute did not show any change. Conclusion: From this study it may be concluded that for interpretation of NESTROFT reading at 5-minute would be optimum. BANGLADESH J CHILD HEALTH 2022; VOL 46 (3) : 130-137

HBB Gene Mutations and Their Pathological Impacts on HbE/β-Thalassaemia in Kuala Terengganu, Malaysia.

β-thalassaemia is a disorder caused by mutations in the β-globin gene, leading to defective production of haemoglobins (Hb) and red blood cells (RBCs). It is characterised by anaemia, ineffective erythropoiesis, and iron overload. Patients with severe β-thalassaemia require lifelong blood transfusions. Haemoglobin E beta-thalassaemia (HbE/β-thalassaemia) is a severe form of β-thalassaemia in Asian countries. More than 200 alleles have been recognised in the β-globin region. Different geographical regions show different frequencies of allelic characteristics. In this study, the spectrum of β-thalassaemia (β-thal) alleles and their correlation with iron overload, in HbE/β-thalassaemia patients, β-thalassaemia trait, and HbE trait were studied. Blood samples (n = 260) were collected from 65 β-thalassaemia patients, 65 parents (fathers and/or mothers) and 130 healthy control individuals. Haematological analyses, iron profiles, and serum hepcidin levels were examined for all participants. DNA was extracted from patients' and their parents' blood samples, then subjected to PCR amplification. Multiplex amplification refractory mutation system PCR (MARMS-PCR) was conducted for eighteen primers to detect the mutations. There was severe anaemia present in HbE/β-thalassaemia patients compared to their parents and healthy controls. The ferritin and iron levels were significantly increased in patients compared to their parents and healthy controls (p = 0.001). Two common mutations were detected among the patient group and three mutations were detected among their parents, in addition to seven novel mutations in HbE/β-thalassaemia patients (explained in results). Some mutations were associated with severe anaemia in β-thalassaemia patients. The detection of mutations is a prognostic marker, and could enhance the appropriate management protocols and improve the haematological and biochemical statuses of β-thalassaemia patients.

PATTERNS AND DEMOGRAPHIC DISTRIBUTION OF HEMOGLOBINOPATHIES DIAGNOSED ON HPLC IN A TERTIARY CARE HOSPITAL OF NORTH EASTERN INDIA

Introduction: Hemoglobinopathies are the most common genetic disorders reported in the World. According to WHO, 5% of the world population is a carrier for hemoglobin disorder. The aim of the study was to determine the Aims and objective: prevalence and demographic prole of hemoglobinopathies in a tertiarty care hospital. During 1 year period, a cross-secti Materials and Methods: onal study was carried out on 150 microcytic hypochromic anaemic patients samples (Hb <11g/dl) of age 1-18 yrs. All the samples were tested for complete blood count and sent for HPLC study. Out of 150 cases, 114 cases were diagnosed as having hemoglobinopathies Results: by HPLC with its prevalence being 76%. With a slight male preponderance was seen (52.6%). The most common hemoglobinoapthy being detected was HbE trait (28%) followed by HbE disease (26.67%). While β thalassemia trait was 13.33%, β thalassemia major was 0.67% and HbE with β thalassemia was 7.33%. The most affected community are the Tribal community with a prevalence of 21.05% followed by the Tai Ahom community (17.54%). Assam is a rich Conclusion: reservoir of hemoglobinopathies as nearly 60% of all cases of anemia coming for investigations may show some form of this disorder. Since there is no effective treatment for this disease, so carrier detection and awareness programmes have to be conducted at community level for its control and prevention

Pattern of hemoglobinopathy among the young tribes of West Bengal: A completely different scenario from Rest of India.

Hemoglobinopathy is a major concern among the tribal population which constitutes 8.6% of the total population, and West Bengal (WB) is the home to 5.3 million tribes. The present study was conducted on 52,880 tribal school students from all the districts of WB. Written informed consent and peripheral blood were collected for complete blood count and high-performance liquid chromatography analysis. Beta trait was 5.3%, sickle trait was 2.35%, and hemoglobin (Hb) E (HbE) trait was 1.4% in this population. About 37.8% of beta trait belonged to the Santal tribe and 21.5% belonged to Oraon. HbS is mainly found in Alipurduar and Jalpaiguri districts at the prevalence of 3.69% and 5.96%, respectively. HbE trait is found at 6.06% in Alipurduar, of which 51% of cases are from Mech tribe only found in this district. Unlike central and Western parts of India, HbS trait in WB was significantly low among the tribes. A high prevalence of consanguinity among the tribes is considered responsible for the high rate of hemoglobinopathy.

Prevalence and spectrum of haemoglobinopathies in females of reproductive age group- A first tertiary care center experience in Punjab, North India.

Thalassemia and hemoglobinopathies are a group of inherited conditions characterized by abnormalities in the synthesis or structure of hemoglobin (Hb). According to estimates, approximately 7% of the world population is a carrier of Hb disorders, leading to high morbidity and mortality. To reduce the burden of these highly prevalent monogenic disorders, detecting them in the carrier stage is crucial to prevent disease progression. We aimed to estimate the prevalence and spectrum of hemoglobinopathies in females in the reproductive (20-40 years) age group. It was a retrospective observational study carried out for 2.5 years (from January 2018 till June 2020). All the females in the age group of 20-40 years age whose blood samples were received in the department for High-Performance Liquid Chromatography (HPLC) were included. The cases with abnormal HPLC findings were analyzed for hematological parameters including hemoglobin, RBC count, and RBC indices [mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), & red cell distribution width - coefficient of variation (RDW-CV)]. Statistical package for social science (SPSS) statistics 21 version for Microsoft Windows (Chicago, USA) was used for statistical analysis of data. The data were described in terms of range, mean ± standard deviation (SD), frequencies (number of cases), and relative frequencies (percentage) as appropriate. During the study period, 72.2% of the females were affected with β-thalassemia trait, followed by HbD Punjab trait (17.8%), HbQ India trait (2.9%), β-thalassemia major (1.8%), and two cases (1.2%) each of HbS trait, HbD Iran trait, and compound heterozygous of HbD Punjab and β-thalassaemia, whereas HbE trait, compound heterozygous of HbQ and β-thalassemia, compound heterozygous of HbJ-variant and β-thalassemia had one case each (0.6%). Preventive strategies are cost-effective and include population screening, premarital screening, screening of spouses, genetic counseling, and prenatal diagnosis. Educating the carrier females about the potential risk and various screening methods may help in controlling the disease.

Hematological Parameters: Manual vs Automated Method; Among the β-Thalassemia and other Haemoglobinopathies in a Tertiary Care Hospital in Kolkata

Background: According to a report of WHO in 2007, 7% of world populations are carrier for Haemoglobindisorder and accurate and timely detection of various Hb variants including beta thalassemia trait can preventoccurrence of more serious disorders like thalassemia major in new-borns. But in a developing country like ususe of high pressure liquid chromatography (HPLC) is limited, manual testing is done and accurate assessmentis quite impossible. So, an observational cross-sectional study was done among 117 β-thalassemia and otherHemoglobinopathies cases and carriers.Objective: To find out the variation between the results of haematological tests obtained by automated countingchamber and manual method.Materials & Methods: This Observational, cross-sectional study was done at Thalassemia control unit and Physiologydepartment of R G Kar Medical College and Hospital, Kolkata. β-thalassemia and other Hemoglobinopathies casesand carriers detected by complete blood count with HPLC from Thalassemia Outpatient Department (OPD) andantenatal mothers from antenatal clinic (ANC) were the population of this study. Findings and variations of thefeatures of different hematological parameters amongst the β-thalassemia and other hemoglobinopathies werereviewed. A finding by HPLC was HbA, HbA2, HbF, HbD, and HbE.Results: Result of only Hb% obtained from HPLC and manual method was significantly different inβ- THALASSEMIA Trait (p=0.0001), HbE Trait (p=0.0001), HbS trait (p=0.0001) and not significant in HbE diseaseand no other Red cell indices were significantly different in two methods. Conclusion: As hematological featureslike TC of RBC or PCV results by manual method shows no significant differences with HPLC/Automated counter assessment, such methods can be utilized even at primary level cost effectively for assessment of hematologicaldisorders. But significant difference is seen in estimating Hemoglobin percentage in between Shalli‘s method andHPLC/Automated counter assessment. So some alternate method may introduce at lower level of health systemother than Shalli’s method

STUDY OF SPECTRUM OF HEMOGLOBINOPATHIES IN ADULT AGE GROUP DIAGNOSED ON HIGH PERFORMANCE LIQUID CHROMATOGRAPHY IN TERTIARY CARE HOSPITAL

Background: Inherited abnormalities of hemoglobin synthesis (hemoglobinopathies) include a myriad of disorders ranging from thalassemia syndromes to structurally abnormal hemoglobin variants. Aims and Objectives: To study spectrum of hemoglobinopathies in adult age group diagnosed on HPLC in tertiary care hospital. Materials and Methods: The present study carried out in the deparment of pathology , DR PDMMC AMRAVATI over a period of 18 months from january 2021 to june 2022. A total of 800 cases were included in the present study. The age group of patients ranged from 19 years and above. HPLC , complete blood count, and peripheral smear examination were done in all the cases. Results and Discussion: In the present study total 800 patients were included out of which 134 cases displayed abnormal hemoglobin fractions on HPLC. It was seen that majority of the patients in study were in the 19yrs to 30yrs. The most common hemoglobinopathy observed in the present study was sickle cell trait (51.50 %) followed by Beta Thalassemia trait (26.12 %) and Sickle cell disease (16.42 %).HbDpunjab trait was observed in 3.74 % cases. Sickle cell Disease + Beta thalassemia trait ,HbE trait, HbE homozygous were observed in 0.74% cases each. Conclusion: Detection of β-thalassemia carriers and other structural variants will provide a preliminary insight to the prevalence of hemoglobinopathies in hospital based patients.Awareness can be created in an effort to prevent the birth of affected children.The present study is intended to contribute to the available data as the identification of these disorders is important epidemiologically and aids in prevention of more serious hemoglobin disorders.

Pregnancy outcomes among women affected with thalassemia traits

ObjectiveTo compare the maternal and perinatal outcomes between a group of pregnant women diagnosed with thalassemia traits and normal controls.Study designA retrospective cohort study was conducted on singleton pregnant women affected and unaffected by thalassemia traits who attended an antenatal care clinic and delivered in Siriraj Hospital. Thalassemia status for all subjects was diagnosed by hemoglobin typing and/or DNA analysis. Patient charts were reviewed from January 2007 to December 2018. The control participants were randomly selected from the same period, with a control–case ratio of around 1:1.ResultsOverall, 1288 women with thalassemia traits (348 with α thal-1 trait, 424 with β thal trait and 516 with HbE trait) and 1305 women in the control group were recruited. Baseline characteristics of both groups were similar, with the exception that the hematocrit level in the first trimester in the thalassemia trait group was significantly lower than that in the control group (34.8 ± 3.4% VS 36.9 ± 3.0%; p < 0.001). The prevalence of pregnancy-induced hypertension (PIH) was higher in the thalassemia trait group, at 6.9% VS 4.7% in the control group; p = 0.018. When subgroups were analyzed between each thalassemia trait, the number of maternal anemias in the first and third trimester was higher for all thalassemia traits compared to the normal group. The β thal and HbE traits increased the risk of PIH, with a relative risk (RR) = 1.67 and 1.66, respectively.ConclusionsThalassemia traits minimally but significantly increase the risk of hypertensive disorders and maternal anemia. In addition, physiological changes during pregnancy may worsen the severity of anemia in the pregnant women with thalassemia traits.

Spectrum of Haemoglobinopathies Detected on Antenatal Screening and Diagnostic Work-up in an Urban Healthcare Set-up: A Retrospective Study

Introduction: Haemoglobinopathies are inherited disorders of haemoglobin (Hb) and consist of Thalassemias and many other structurally variant haemoglobins. Out of these, beta-thalassemia major and clinically significant sickle cell disorders are of great public health importance in India. Lack of awareness regarding their prevalence and knowledge about diagnostic methods, has resulted in failure of community control of birth of these otherwise totally preventable genetic disorders in India. Aim: To study the spectrum of haemoglobinopathies in the study population and to assess the effectiveness of the antenatal screening program, in identifying the couples at-risk and providing prenatal intervention. Materials and Methods: This retrospective cohort study was performed at a community healthcare set-up in Mumbai. from August 2021 to October 2021 on medical records of 10,025 patients including women who were part of antenatal screening, patients investigated for anemia and who underwent haemoglobin electrophoresis over 12 years (October 2007 to October 2019). Alkaline Electrophoresis test was performed using the InterlabGenios analyser on cellulose acetate and the findings were interpreted along with haemogram parameters. Finding of various conditions were presented in terms of prevalence rates and mean values of relevant blood count parameters were presented as mean±SD. Results: An abnormal haemoglobin pattern was seen in 544 (5.42%) of the 10,025 cases with age group ranged from 1 year to 84 years. Most common haemoglobinopathies detected were beta (β) thalassemia trait 378 (3.77%) followed by sickle cell trait (0.9%), Hb D trait (0.26%), HbE trait (0.24%) and others. Carriers of haemoglobinopathies were detected in 186(3.74%) of 4968 women, on antenatal screening. A total of 18 couples-at-risk were identified. One child with thalassemia major and another with sickle cell disease were born in this population over 12 years. Conclusion: The b-Thalassemia trait and HbS trait are the commonest haemoglobinopathies detected. Antenatal screening programme and timely intervention is an effective strategy to control clinically significant major hemoglobinopathies.

Prevalence of Haemoglobin Variants and Haemoglobinopathies in a Single Paediatric Centre in Southern India: A Retrospective Cross-sectional Study

Introduction: Haemoglobinopathies are qualitative disorders of Haemoglobin (Hb) resulting from structural defects in the amino acid sequence of one of the globin chains, whereas, thalassaemia results from quantitative defects in the synthesis of one or more of the globin chain subunits of the Hb tetramer. Cation Exchange- High Performance Liquid Chromatography (CE-HPLC) is one of the methods for initial screening of Hb variants like HbS, HbD, HbE etc and for quantification of HbF, HbA and HbA2 levels. Aim: To find out the prevalence of haemoglobinopathies in patients of a Government paediatric tertiary care hospital in south India. Materials and Methods: In this retrospective, cross-sectional study, all laboratory requests, in the period from August 2019 to July 2021, for Hb variant analysis by HPLC were collected, irrespective of provisional diagnosis. The Hb variant analysis was carried out by CE-HPLC on the Bio-Rad D-10 analyser. This study was conducted in a Government Paediatric tertiary care hospital in Southern India for patients who had any clinical or familial suspicion of haemoglobinopathies. The Statistical analysis was performed using Microsoft Excel 2010. Results: Total data of 704 laboratory requests for Hb Variant analysis were obtained. Out of 704 laboratory request, 585 were from children younger than 12 years of age and 119 were parental screening. There were 164 abnormal chromatograms. Out of 164 patients, 91 were female patients and 73 were male patients. Out of 164 abnormal chromatograms, 97 (59.15%) were beta- thalassaemia trait, 18 (10.98%) were beta-thalassaemia major, 13 (7.93%) were sickle cell trait, 2 (1.22%) were sickle cell disease, 21 (12.80%) were HbE trait, 2 (1.22%) were homozygous HbE, 2 (1.22%) were HbD trait, 3 (1.83%) were Hereditary Persistence of Foetal Hb (HPFH)/delta beta-thalassaemia, 2 (1.22%) were HbJ trait, 1 (0.61%) was HbE beta-thalassaemia, 1 (0.61%) was sickle- beta-thalassaemia and 2 (1.22%) were alpha thalassaemia. Conclusion: From this study, beta-thalassaemia trait and beta- thalassaemia major were found to be the first and second most prevalent haemoglobinopathies in children below 12 years of age. This data suggests the importance of premarital and antenatal screening procedures that can help in reducing the possibility of such haemoglobinopathies in the future generation, suffering and burden of disease to the family and society.

Risk Factors Associated with Diabetic Retinopathy and Thalassemia Prevalence Survey in Diabetes Mellitus Patients at Six Primary Care Units of Naresuan University Hospital: A Cross-sectional Study

Objective: To evaluate the risk factors and prevalence of diabetic retinopathy (DR) in both medical and socioeconomic aspects and find prevalence of thalassemia which associated hemoglobin A1c (HbA1c) measurement in diabetes mellitus (DM) patients at six primary care units (PCU) of Naresuan University Hospital (NUH). Materials and Methods: A cross-sectional survey of DM patients participated in annual proactive DR screening program at six PCU of NUH between December 2016 and March 2017 was conducted. Medical data were retrieved from medical records at PCU. Patients were also interviewed to gather socioeconomic information. Fundus examination was done by indirect ophthalmoscope. Three milliliters of blood was collected from each patient on the same day for Hb analysis. Results: Four hundred and eighty-eight DM patients participated in the present study. Mean age, duration of DM, fasting blood sugar (FBS) level, and HbA1c level were 61.2±9.8 years, 8 years (4 to 12), 124 mg/dL (108 to 151.5), and 7.1% (6.5 to 8.1), respectively. Prevalence of overall DR was 2.9% (14 patients) and proliferative DR was 0.2% (1 patient). Risk factors of DR were HbA1c at 7% or more [adjusted OR 4.7 (95% CI 1.4 to 13.5) and p=0.011] and emotional stress [adjusted OR 3.3 (95% CI 1.1 to 9.8) and p=0.033). Thalassemia screening found 116 patients had abnormal hemoglobin. Ninety-three patients were HbE trait, eight were HbE, ten were alpha-thalassemia trait, two were beta-thalassemia trait, one was HbH, one was alpha- and beta-thalassemia trait (α/β), and one was alpha-thalassemia trait and HbE trait (α/E), and all of them were thalassemia minor or intermedia. Only four patients from HbE trait group had DR. The mean HbA1c in all groups of patients with either normal or abnormal hemoglobin were not statistically significant different. Conclusion: The present study showed that HbA1c and emotional stress might have played an important role in association with DR development. Thalassemia minor and intermedia seemed not to associate with HbA1c measurement. Keywords: Diabetic retinopathy; Thalassemia; Primary care unit; Naresuan university; Risk factors

Distribution of β-Thalassemia and Other Hemoglobinopathies in Bangladeshi University Students and Ready-Made Garment Workers.

BackgroundThe inheritable thalassemia and other structural hemoglobinopathies are prevalent globally. In Bangladesh, there is a scarcity of studies concerning thalassemia and other structural hemoglobinopathies. Hence, the present study investigated the prevalence and distribution of β-thalassemia and other hemoglobinopathies among the university students and ready-made garment (RMG) workers.MethodsA cross-sectional study was conducted, whereas a total of 989 subjects were selected randomly from 10 garments industries (n=495) and five universities (n=494) for diagnosis and counseling from April 2018 to August 2018. Basic information related to the study and venous blood was collected and subjected to capillary Hb-electrophoresis for each participant.ResultsThe present study identified thalassemia carriers and other hemoglobinopathies among 13.6% of the participants (14.1% university students and 13.2% RMG workers). The most common form of hemoglobin formation disorder was the Hb-E trait (10.5%), considering both participants’ groups. Other forms of hemoglobin formation disorders observed were a β-thalassemia trait, Hb-D trait, Hb-E disease, and β-thalassemia major. The hemoglobinopathies were significantly associated with the individuals who had a family history of thalassemia and were not vaccinated against hepatitis B in the total sample. Although not statistically suggestive, the most considerable portion (87.3%) of the carriers was unmarried in the present findings.ConclusionThe present study findings are anticipated to help the public health authorities by providing novel data (of a neglected cohort, ie, RGM workers), facilitating the upcoming comprehensive national thalassemia prevention programs.

THALASSEMIA SYNDROME AND OTHER HEMOGLOBINOPATHIES: A SINGLE THALASSEMIA CENTRE EXPERIENCE FROM TERTIARY CARE HOSPITAL OF PURULIA

Background: Hemoglobinopathies are one of the major health burden globally including India and Southeast Asian region. This study was carried out to nd out the prevalence of β-thalassemia, HbS, HbD, HbE as well as identication of asymptomatic carriers who have an increased risk of having a child with thalassemia in the western part of West Bengal. Materials and methods: In this retrospective cohort study, the data was collected from all newly registered patients with anemia, referred from different outdoor patients in a tertiary care hospital, attending thalassemia clinic over a period of 3 years from January2018 to December2020. Detailed clinical history was taken and blood samples collected are tested with Sysmex automated blood cell counter for red cell indices. Diagnosis of hemoglobinopathy was done by G8 HPLC (high performance liquid chromatography) Analyzer by TOSOH Bioscience. Results: A total 2297 cases were studied during the study period. The age of the patients ranged between 6 months to 60 years. Higher percentage of the study sample was at age of more than 10 years (93.8%). Among 2297 cases, 477 cases showed abnormal Hb fractions on HPLC. The following Hb abnormalities detected were following: β (beta) thalassemia trait 13.2%, β thalassemia major 2.3% followed by sickle cell trait 2.17%, HbE trait 1.8%, HbS disease 0.21%, Hb E disease 0.13%, hereditary persistence of fetal hemoglobin (HPFH) traits 0.17% and Hb D trait 0.08% along with HbE-β-thalassemia 0.30% and HbS-β-thalassemia 0.26%. Conclusion: Among the hemoglobinopathies, β-thalassemia trait (12.9%) is prevalent in western part of Bengal.

489 Pregnancy outcomes among women affected by thalassemia traits compared to normal controls

To compare maternal and perinatal outcomes between groups of pregnant women diagnosed with thalassemia traits and normal controls. A retrospective cohort study was conducted on singleton pregnant women affected and not affected by thalassemia traits who attended in antenatal care clinic and delivered in Siriraj Hospital, Thailand. Thalassemia status for all subjects was indicated by hemoglobin typing and/or DNA analysis. Patients’ charts from January 2007 to December 2018 were reviewed. The control participants were randomly selected in the same period with a control-case ratio of approximately 1:1. Overall, 1288 women with thalassemia traits (348 of α thal-1 trait, 424 of β thal trait and 516 of HbE trait) and 1305 women in the control group were recruited. Maternal age, BMI and parity were similar between two groups. The hematocrit level at the 1st trimester in the thalassemia trait group was significantly lower than that in the control group (34.8±3.4% VS 36.9±3.0%; p<0.001). Table 1; the risk of pregnancy induced hypertension (PIH) and Apgar score < 7 at 1 minute were significantly higher in the thalassemia trait group compared to the control group (6.9% VS 4.7%, p=0.018 and 4.1% VS 2.5%, p=0.02). A subgroup was also analyzed between each thalassemia trait (Table 2), showing that the numbers of maternal anemia in the 1st and 3rd trimester were higher in all thalassemia traits compared to the normal group (p<0.001). β thal trait and HbE trait increased the risk of PIH with relative risk (RR) = 1.67 and 1.66, respectively. Additionally, the rate of neonates with Apgar score < 7 at 1 minute was significantly higher in the HbE trait; RR = 2.06. There was no significant difference in other maternal and neonatal outcomes. Thalassemia traits significantly increase the risk of hypertensive disorders in pregnancy and low Apgar score at 1 minute. Physiologic change during pregnancy may worsen the severity of anemia in pregnant women affected by thalassemia traits.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Evaluation of role of HPLC (Merits & Pitfalls), in the diagnosis of various hemoglobinopathies & thalassemic syndromes.

: HPLC is one of the most important tools for accurate diagnosis of hemoglobinopathies and thalassemias. The advantage of the HPLC system is the excellent resolution, reproducibility &quantification of several normal and abnormal hemoglobin. BIO RAD Variant II analyzer was used. Sickle cell syndromes including double heterozygous states accounted for 56.13% of total cases. HbSS, HbS/β0-th, HbS/β+-th β-thal trait comprises 29%, 6.5%, 5.1%& 10% of total cases respectively with mean MCV (fl) = 84, 68,71,64 respectively. The Mean HbA2 for β-thal trait, HbE trait &HbE-β thal showed 5.1 ± 1.1, 19 ± 9 & 24 ± 8 respectively. HbF is increased in 8.6% case (excluding SC syndromes & β-thal disorders), of these 5.5% were infants & 12 cases of Aplastic Anemias. Peak P2 >7% (2.4% cases) was seen in uncontrolled diabetes mellitus which on quantification showed HbA1C = 8 ± 2.1 mmol/L. : HPLC in correlation with CBC parameters & family studies can aid in the diagnosis of majority of Hemoglobinopathies and thalassemic syndrome. The CBC & HPLC parameters of the present study are in good correlation with the research conducted by Tejinder Sing, RiouJ & Alla Joutovsky. Present study showed HPLC comprehensively characterizing HbS, A, A2, F, S, C, D from each other & was also applicable for the quantification of HbA1c for the monitoring of Diabetes Mellitus. : The merits of HPLC are small quantity of sample required, economical, less TAT, accurate categorization of HbS, HbA2 & F. But one has to be aware of the limitations and problems associated with this method due to variant hemoglobin within the same retention windows. The present findings show HPLC as an excellent & powerful diagnostic tool for the direct identification of hemoglobin variants with a high degree of precision in the quantification of normal and abnormal hemoglobin fractions.

Pattern of hemoglobinopathies and thalassemia in Manipur, India

Background: Hemoglobinopathies are the commonest genetic disorders worldwide. Thalassemia Major, Thalassemia Intermedia and Sickle Cell Disease are the major disorders that require lifelong management and are to be considered for prevention. In India, Beta-Thalassemia is prevalent across the country, with an average frequency of carriers being 3-4%.Methods: This is a cross sectional study conducted between June 2016 - May 2017 in the Department of Medicine, RIMS Imphal in 453 patients as a workup for anemia and clinically suspected cases of Hemoglobinopathy or beta thalassemia. Blood samples were collected and sent for Haemoglobin Electrophoresis using cellulose alkaline technique.Results: Among the 453 cases of the population surveyed, 35% showed the presence of abnormal hemoglobin. 16% were found to be beta thalassemia carrier, 11.69% HbE trait, 6.62% Homozygous HbE, 0.4% beta thalassemia and 0.7% had Hereditary persistence of HbF.Conclusions: High prevalence of Beta Thalassemia trait occurred more frequently than other Hemoglobinopathies. The study concludes that it is immensely important epidemiologically to explore the haemoglobin variants in Manipur so that the carriers can be detected for prevention of more serious disorder in the future generations.

A demographic prevalence of β Thalassemia carrier and other hemoglobinopathies in adolescent of Tharu population.

Background and Aims:Hemoglobinopathies and thalassemias are the commonest single gene disorders in India. In Terai region of India, Hemoglobinopathies and thalassemias are the most common in the Tharu community. Therefore, in this study, we aim to evaluate the Hb variant analysis of hemoglobinopathies and thalassemias in a Tharu population in Lakhimpur Kheri Districts of Uttar Pradesh, India.Materials and Methods:Total 493 individuals were recruited in this study. The demographic details and blood samples were collected from different location at Kheri district during mega health camp. Hb variant analysis was performed by high performance liquid chromatography (HPLC) system beta thalassemia short program in BIO-RAD VARIANT.Results:Out of 493, 108 (21.9%) individual suffers with abnormal haemoglobinopathies. In which β-thalassemia trait is the commonest haemoglobinopathy (12.98%), followed by HbE trait (7.50%), and compound heterozygous HbS/β-Thalassemia trait (1.42%) in overall population. The HbF was significantly greater in HbS heterozygous (1.45 ± 1.41), whereas mean HbA2 was significantly greater in β-Thalassemia trait (5.17 ± 1.36).Conclusion:The high incidence of hemoglobinopathies and thalassemias were observed in Tharu community in Lakhimpur Kheri districts of Uttar Pradesh, Indian.

A "One-Stop" Screening Protocol for Haemoglobinopathy Traits and Iron Deficiency in Sri Lanka.

Introduction: The high frequencies of carriers of severe haemoglobinopathies and of iron deficiency in Southeast Asia require reliable and affordable tests to improve on current screening procedures.Objectives: We evaluate a “one stop” approach using the THALCON dichlorophenolindophenol (DCIP) and one-tube osmotic fragility (OF) tests and measurement of Zinc Protoporphyrin (ZPP) to detect and distinguish HbE and β-thalassaemia traits from iron deficiency. We compare findings with current screening practice in Sri Lanka that relies on the identification of low mean red cell volume and/or mean red cell hemoglobin for this purpose.Methods: Between November 2017 and May 2018, we undertook a cross-sectional survey of secondary school students in Gampaha district, Sri Lanka. The THALCON–DCIP and OF tests were compared to capillary electrophoresis (CE), used as a gold standard to detect haemoglobinopathies. ZPP was measured in whole blood. Plasma ferritin and C-reactive protein (CRP) were measured in students with a raised ZPP concentration.Results: We collected venous blood samples from 1,324/1,332 (99.4%) students. The median age of the students was 17 (IQR 16–18) years, all were Sinhalese and 814/1,324 (61.5%) were female. CE identified 3 students with HbE trait and 26 students with β-thalassaemia trait. The THALCON–DCIP test was positive only in the 3 students with HbE (sensitivity 100%, 95% CI 29.2–100.0; specificity 100%, 95% CI 99.7–100.0). The THALCON–OF test identified all 26 students with β-thalassaemia trait (sensitivity = 100%, 95% CI 86.8–100.0) and 287 students with a normal CE result (specificity = 77.9%; 95% CI 75.5–80.1). It was also positive in 2/3 (66.7%) students with HbE trait. Iron deficiency (ZPP > 70 μmol/mol heme) was present in 118/1,240 (9.5%) students with a normal hemoglobin genotype, all of whom had plasma ferritin <15 ng/ml and CRP <5 mg/L.Conclusion: This one–stop approach offers reliable and affordable population screening for both haemoglobinopathy traits and iron deficiency in resource-limited settings where these conditions are common and ensures that iron supplements are targeted only to those who require them. Further work is warranted to refine the OF test to reduce the number of false positive results.

Clinical Profile of Thalassemia Syndrome in Children of Northern Bangladesh

Background: Thalassemia is a common hematological disorder in our country having wide spectrum of clinical presentation. The frequency and severity of the several types of thalassemia depend on the racial background of the population. Hb-E Beta thalassemia is prevalent in our country.&#x0D; Objective: To see the clinical features of different types of Thalassemia in northern area of Bangladesh. Methods: Hundred cases were selected from Thalassemia patient admitted in department of pediatrics, on May 2012 to October 2012. A prescribed questionnaire was used to record the information. The methods were explained to the patients and consent was taken. Necessary physical examination was performed and investigations were done. The data was analyzed by standard procedure.&#x0D; Results: Out of hundred (100) cases, most (61%) were Hb-E beta Thalassemia, less common (1%) was Hb-E disease, and 1 % case was Hb-E trait. Majority (64%) manifested clinically under one year of age. 54% were male and 46% were female. The major presenting symptom was progressive pallor in 70% cases. Others presenting complaints were low grade fever (40%). Hemoglobin concentration at the time of diagnosis was below 5 gm/dl in 53.33% patients. In hemoglobin electrophoresis it was Hb-E ranged from 54.64 ± 13.02%, Hb-F 34.84±13.73%, Hb-A 23.32± 18.15% and Hb-A2 3.5± 70%. Radiological findings revealed gross bony changes occur in long standing cases. Enlarged cardiac shadow was found in those cases having severe anemia with heart failure.&#x0D; Conclusion: In countries with a high incidence of thalassemia, it is vitally important to offer prospective genetic counseling and to warn carriers about the risks of intramarriage. Nutritional and folic acid supplementation with regular blood transfusion along with iron chelation therapy is essential to improve the prognosis.&#x0D; TAJ 2018; 31(2): 6-11

The Importance of Prenatal Diagnosis By Chorionic Villous Sampling in Detecting and Preventing Beta Haemoglobinopathies in a Resource Poor Setting: A Report from Eastern Part of India

Introduction: β Haemoglobinopathy, is one of the commonest inherited disorders, with a higher prevalence in the eastern part of India. In view of the lack of facilities for allogeneic stem cell transplantation, including financial constraints, this disease is considered to be incurable in this part of India. So, preventing the birth of a symptomatic Thalassemic child is the best way to prevent the disease, considering the immense medical, social & financial implications in the community. Therefore, prenatal diagnosis is the most important tool for detecting & preventing symptomatic haemoglobinopathies.Objectives: To detect the status of haemoglobinopathies in the foetus of carrier or affected parents, so as to prevent the birth of symptomatic patients with Thalassemia & other haemoglobinopathies.Material & Methods: This is a retrospective analysis of an ongoing registry data of routine prenatal diagnosis of haemoglobinopathy, conducted at NRS Medical College, Kolkata, over a period of approximately 4 ½ years (September,2013-April,2018). The process involves, collection of Ultrasound guided chorionic villous samples (CVS) by an Obstetrician between 12-16 weeks of gestation (maximum up to 19 weeks). Considering the prevalence of specific mutations in this part of India, a total of 25 β mutations are analysed by ARMS-PCR, using specific probes. Some of the common mutations tested are: IVS1-5(G→C), 619bp del, IVS 1-1(G→T), CD 30(G→C), CD 26(G→A) & CD6 (A→T).Results: Total number of CVS were 407(407 mothers were tested), of which 329 samples (80.8%) showed presence of β haemoglobinopathy mutations, & 78(19.2%) did not have any mutations. Among the mutated genotype, heterozygous carrier state was detected in 232(57%), & homozygous/ compound heterozygous state(affected) was detected in 92 foetuses (22.6%). Among the carriers, β Thalassaemia trait was most common(n=185,45.45%), followed by HbE Trait(n=41,10.07%). HbS trait(n=3), HbD trait(n=1) & δβ Thalassemia trait (n=2) were rarely detected. Among the affected foetuses, 44 had compound heterozygous genotype (10.8%), majority being HbEβ thalassemia (n=42,10.3%), followed by δβ-β Thalassemia(n=1) & HbSβ Thalassemia(n=1). The remaining 48 affected cases were homozygous β Thalassemia Major (11.7%). Of the 92 affected pregnancies, 91(22.35%) had been counselled for medical termination of pregnancy (MTP). One case of δβ-β Thalassemia, a milder phenotype, was excluded from MTP counselling. Total number of β mutations detected (both homozygous & heterozygous states) were n=373, and the commonest was IVS1-5(G→C) (n=244,65.4%), followed by CD 26(G→A) (n=85,22.78%), CD 30(G→C) (n=9,2.4%), CD 41/42(-CTTT) (n=8,2.1%), CD 15(G→A) (n=8,2.1%) & CD8/9 (+G) (n=4,1.1%). Rarer β mutations detected were CD15(-T) (n=4,1.1%), CD16(-C) (n=1), CD6 (A→T) (n=4,1.1%), βD-PunjabCD121 (G→C) (n=3) & Asian Indian inversion deletion Gγ (Aγδβ)0(n=2).We know that sometimes maternal cell contamination may occur during villous sampling. So, at the time of childbirth, cord blood was collected & β mutation studies were re-performed. If this could not be done, an HPLC was conducted when the child was >1year old. We have data of 26 children & each time the results have corroborated with the primary CVS sampling data.Among both male and female parents(n=814), the commonest genotype was β Thalassemia Trait (n=621, 76.3%). Among the female parents(n=407), some of the other genotypes were HbE Trait (n=94,23.09%) & HbS trait(n=4,0.98%). Among the male parents(n=407), HbE Trait(n=70,17.19%) was the 2nd most common abnormality, few of the other genotypes being: HbS Trait (5,1.2%), β Thalassemia major (n=3,0.73%) & HbE homozygous disease (n=3, 0.73%).Conclusions: From this cohort, total affected (homozygous/compound heterozygous) foetuses are 92(22.6%). β Thalassemia Major (11.7%) is the most common affliction, followed by HbEβ thalassemia (10.3%) & δβ - β Thalassemia. An attempt at prevention of birth of at least 91(22.35%) future thalassemia patients has been done by means of MTP.Ours is an ongoing registry data and we are following up the children by conducting cord blood mutation studies or HPLC (>1year old child), to study the precision and accuracy of our CVS sampling results. So far there has been 100% concordance. One of the limitations of our study is lesser number of reconfirmation of the born children, in view of social and financial constraints. DisclosuresNo relevant conflicts of interest to declare.