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BACKGROUND AND AIMS: To update the molecular characteristics of α-thalassemia in northeast Thailand, the molecular basis and genetic interactions of Hb H disease were examined in a large cohort of patients. MATERIALS AND METHODS: A study was done on 1,170 subjects with Hb H disease and various genetic interactions encountered during 2009-2023. Hb and DNA analyses were carried out. RESULTS: As many as 40 genotypes with several known, previously undescribed, and novel mutations were observed. These included 698 subjects (59.8 %) of Hb H disease, 357 (30.6 %) with EABart's disease, 63 (5.4 %) with EEBart's disease, 18 (1.7 %) with abnormal Hbs, 17 (1.5 %) with ß-thalassemia, and 4 (0.4 %) with EFBart's or EFABart's disease. The molecular basis of 13 subjects (1.1 %) remains unknown. The α0-thalassemia included --SEA (n = 1,139, 97.4 %) and --THAI (n = 21, 1.8 %). Two rare mutations were identified in 3 subjects (0.3 %) with --SA and --CR deletions. For α+-thalassemia, -α3.7 kb del (n = 626, 53.5 %), Hb Constant Spring (n = 415, 35.5 %), -α4.2 kb del (n = 44, 3.8 %), Hb Paksé (n = 36, 3.1 %), and Hb Q-Thailand (n = 19, 1.6 %), were detected. Ten rarer α+-thalassemia were identified, including a novel mutation, namely the Hb Chumphae (HBA2:c.32T>A). The Hb H-Lansing-Ramathibodi, Hb H-Jax, and Hb H-Chumphae are hitherto undescribed in this region. PCR-based diagnostic methods for these α-thalassemia defects were described. CONCLUSIONS: This study confirms the diverse heterogeneity and genetic interactions causing Hb H disease in northeast Thailand. The results should prove useful for laboratory diagnosis and genetic counseling of this genetic disorder in the region.
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Mutação , Talassemia alfa , Humanos , Talassemia alfa/genética , Talassemia alfa/diagnóstico , Tailândia , Estudos de Coortes , Masculino , Feminino , Hemoglobina H/genética , Genótipo , Adulto , AdolescenteRESUMO
BACKGROUND AND AIMS: High hemoglobin F determinants can be classified into hereditary persistence of fetal hemoglobin (HPFH) and δß-thalassemia with different phenotype. We report the molecular basis and hematological features in a large cohort of deletional high Hb F determinants in Thailand. MATERIALS AND METHODS: Subjects (n = 28,177) encountered during 2015-2022 were reviewed, and those with phenotypically suspected of having high Hb F determinants were selected. Combined PCR, multiplex ligation-dependent probe amplification, next-generation sequencing, and DNA sequencing were used to identify the mutations. RESULTS: Among 28,177 subjects investigated, 300 (1.06 %) were found to carry deletional high Hb F determinants in a total of 302 alleles, including heterozygote, compound heterozygote with ß-hemoglobinopathies, and homozygote. DNA analysis identified eight different DNA deletions, including δß0-thalassemia (12.6 kb deletion) (73.8 %), HPFH-6 (14.9 %), Indian deletion-inversion Aγδß0-thalassemia (3.6 %), Thai deletion-inversion-insertion Aγδß0-thalassemia (3.0 %), SEA-HPFH (3.0 %), Chinese Aγδß0-thalassemia (1.0 %), Thai δß0-thalassemia (11.3 kb deletion) (0.3 %), and a novel δß0-thalassemia (137.1 kb deletion) (0.3 %). In addition, three novel genetic interactions, including Chinese Aγδß0-thalassemia/Hb E, δß0-thalassemia/Indian deletion-inversion Aγδß0-thalassemia, and homozygous δß0-thalassemia were found. Hematological features and Hb analysis results of 20 different genotypes were recorded. Multiplex gap-PCR assays for detection of these genetic determinants were described. CONCLUSIONS: Deletional high Hb F determinants are common and heterogeneous in Thailand. Data on the prevalence, molecular spectrum, phenotypic expression, and complex interactions of these genetic determinants should prove useful in the study and a prevention and control program of hemoglobinopathies in the region.
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Hemoglobinopatias , Talassemia , Talassemia beta , Humanos , Hemoglobina Fetal/genética , Tailândia , Talassemia beta/diagnóstico , Mutação , Reação em Cadeia da Polimerase Multiplex , DNARESUMO
AIMS: Krüppel-like factor 1 (KLF1) is an erythroid-specific transcription factor playing an important role in erythropoiesis and haemoglobin (Hb) switching. Biallelic KLF1 mutations can cause haemolytic anaemia with thalassaemia-like syndromes but are rarely reported. We explore the KLF1 mutations in Thai subjects with unexplainable haemolytic anaemia. METHODS: The study was done on 57 subjects presented with haemolytic anaemia and elevated Hb F without ß-thalassaemia diseases. Hb analysis was performed using capillary electrophoresis. Analyses of α-thalassaemia, ß-thalassaemia and KLF1 genes were performed using PCR-based methods and DNA sequencing. RESULTS: Thirteen subjects with compound heterozygous for a known and five new genetic KLF1 interactions were identified, including KLF1:c.519_525dupCGGCGCC/c.892G>C with class 3/2 (n=8), and each subject with new genetic interaction, including KLF1:c.-154C>T;643C>T/c.983G>A with class 3/2, KLF1:c.-154C>T;643C>T/c.809C>G with class 3/2, KLF1:c892G>C/c.983G>A with class 2/2, KLF1:c.892G>C/c.1001C>G with class 2/2 and KLF1:c.1001C>G/c.1003G>A with class 2/2. Most of them had anaemia with Hb levels ranging from 45 to 110 g/L, hypochromic microcytosis, aniso-poikilocytosis, increased Hb F levels (17.9%-47.4%), small amounts of Hb Bart's, regular blood transfusion, hyperbilirubinaemia, increased serum ferritin and nucleated red blood cell. CONCLUSIONS: Biallelic KLF1 mutations associated with anaemia may not be uncommon in Thailand. Characteristics of haemolytic anaemia, abnormal red cell morphology with nucleated red blood cells and elevated Hb F, and presenting small amounts of Hb Bart's without thalassaemia diseases are useful markers to further investigation of the KLF1 gene.
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High Hb F determinants are genetic defects associated with increased expression of hemoglobin F in adult life, classified as deletional and non-deletional forms. We report the first description of non-deletional hereditary persistence of fetal hemoglobin (HFPH) in Thailand. Study was done on 388 subjects suspected of non-deletional HPFH with elevated Hb F expression. Mutations in the Gγ- and Aγ-globin genes were examined by DNA analysis and rapid diagnosis of HPFH mutations were developed by PCR-based methods. Twenty subjects with five different mutations were identified including three known mutations, - 202 Aγ (C>T) (n = 3), - 196 Aγ (C>T) (n = 3), and - 158 Aγ (C>T) (n = 12), and two novel mutations, - 117 Aγ (G>C) (n = 1) and - 530 Gγ (A>G) (n = 1). Interaction of the - 117 Aγ (G>C) and Hb E (HBB:c.79G>A) resulted in elevation of Hb F to the level of 13.5%. Two plain heterozygous subjects with - 530 Gγ (A>G) had marginally elevated Hb F with 1.9% and 3.0%, whereas the proband with homozygous - 530 Gγ (A>G) had elevated Hb F of 11.5%. Functional prediction indicated that the - 117 Aγ (G>C) and - 530 Gγ (A>G) mutations dramatically alter the binding of transcription factors to respective γ-globin gene promotors, especially the CCAAT and GATA-1 transcription factors. Diverse heterogeneity of non-deletional HFPH with both known and new mutations, and complex interactions of them with other forms of thalassemia are encountered in Thai population.
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Fatores de Transcrição GATA , gama-Globinas , Adulto , Humanos , Tailândia , Sítios de Ligação , Fatores de Transcrição , Hemoglobina Fetal , MutaçãoRESUMO
Background: ß 0-thalassemia deletion removing 5´ß-globin promoter usually presents phenotype with high hemoglobin (Hb) A2 and Hb F levels. We report the molecular characteristics and phenotype-genotype correlation in a large cohort of the ß 0-thalassemia with 3.4 kb deletion. Methods: A total of 148 subjects, including 127 heterozygotes, 20 Hb E-ß-thalassemia patients, and a double heterozygote with α-globin gene triplication, were recruited. Hb and DNA analysis were performed to identify thalassemia mutations and four high Hb F single nucleotide polymorphisms (SNPs) including four base pair deletion (-AGCA) at A γ-globin promoter, rs5006884 on OR51B6 gene, -158 G γ-XmnI, BCL11A binding motifs (TGGTCA) between 3´A γ-globin gene and 5´Î´-globin gene. Results: It was found that heterozygous ß 0-thalassemia and Hb E-ß 0-thalassemia with 3.4 kb deletion had significantly higher Hb, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin and Hb F values as compared with those with other mutations. Co-inheritance of heterozygous ß 0-thalassemia with 3.4 kb deletion and α-thalassemia was associated with even higher MCV and MCH values. The Hb E-ß 0-thalassemia patients carried a non-transfusion-dependent thalassemia phenotype with an average Hb of around 10 g/dL without blood transfusion. A hitherto undescribed double heterozygous ß 0-thalassemia with 3.4 kb deletion and α-globin gene triplication presented as a plain ß-thalassemia trait. Most of the subjects had wild-type sequences for the four high Hb F SNPs examined. No significant difference in Hb F was observed between those of subjects with and without these SNPs. Removal of the 5´ß-globin promoter may likely be responsible for this unusual phenotype. Conclusions: The results indicate that ß 0-thalassemia with 3.4 kb deletion is a mild ß-thalassemia allele. This information should be provided at genetic counseling and prenatal thalassemia diagnosis.
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Talassemia beta , Humanos , Talassemia beta/diagnóstico , gama-Globinas , Genes Reguladores , Hemoglobina A2 , Heterozigoto , Estudos RetrospectivosRESUMO
OBJECTIVE: To determine the frequency and etiology of unnecessary prenatal diagnosis for hemoglobinopathies during 12 years of services at a single university center in Thailand. METHODS: We conducted a retrospective cohort analysis of prenatal diagnosis during 2009-2021. A total of 4,932 couples at risk and 4,946 fetal specimens, including fetal blood (5.6%), amniotic fluid (92.3%), and chorionic villus samples (2.2%) were analyzed. Identification of mutations causing hemoglobinopathies was carried out by PCR-based methods. Maternal contamination was monitored by analysis of the D1S80 VNTR locus. RESULTS: Among 4,946 fetal specimens, 12 were excluded because of poor PCR amplification, maternal contamination, non-paternity, and inconsistency of the results of the fetuses and parents. Breakdown of 4,934 fetuses revealed 3,880 (78.6%) at risk for the three severe thalassemia diseases, including ß-thalassemia major, Hb E-ß-thalassemia, and homozygous α0-thalassemia, 58 (1.2%) at risk for other α-thalassemia diseases, 168 (3.4%) at risk for ß+-thalassemia, 109 (2.2%) at risk for high Hb F determinants, 16 (0.3%) at risk for abnormal Hbs, and 294 (6.0%) with no risk of having severe hemoglobinopathies. The parents of 409 (8.3%) fetuses had inadequate data for fetal risk assessment. Overall, we encountered unnecessary prenatal diagnostic requests for 645 (13.1%) fetuses. CONCLUSIONS: The frequency of unnecessary prenatal diagnosis was high. This could lead to unnecessary risk of complications associated with fetal specimen collection, psychological impacts to the pregnant women and their families, as well as laboratory expenses and workload.
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Hemoglobinopatias , Talassemia alfa , Talassemia beta , Gravidez , Feminino , Humanos , Estudos Retrospectivos , Diagnóstico Pré-Natal/métodos , Hemoglobinopatias/diagnóstico , Hemoglobinopatias/epidemiologia , Hemoglobinopatias/genética , Talassemia beta/diagnóstico , Talassemia beta/epidemiologia , Talassemia beta/genética , Talassemia alfa/genética , Líquido AmnióticoRESUMO
BACKGROUND: The wide variation in hemoglobin (Hb) F levels has been observed in patients with Hb EE disease. This study aimed to describe hematologic features and determine the effect of genetic variants on Hb F expression in young children with Hb EE disease. METHODS: Hematologic features and Hb profiles of Laotian children aged 6-23 months, who originally enrolled in the Lao-Zinc study, were retrospectively reviewed. Only children with Hb EE disease, as indicated by DNA analysis, were included in this current analysis. Genetic variants, including the G γ-XmnI polymorphism (C>T) of the HBG2 gene, the HBS1L-MYB intergenic region on chromosome 6, and the BCL11A on chromosome 2 as well as the mutations occurring on the Krüppel-like factor 1 (KLF1) gene, were examined. RESULTS: In total, 205 children were diagnosed as having Hb EE disease with Hb F ranged from 1.2 to 43.7%. Most of the children had mild to moderate anemia with a remarkable microcytosis. Analysis of the genetic variants revealed an extremely high frequency of the G γ-XmnI (93.7%). Applying multiple regression analysis adjusted for age, sex, and α-thal gene, a positive relation was observed for the rs4671393 (coefficient = 3.87, p = .005) and the rs2297339 (coefficient = 2.48, p = .046), but not the G γ-XmnI. A statistically non-significant relation was noted for the rs9399137 and the -154 (C>T) KLF1 mutation. CONCLUSION: Our findings provide insight into complex situation of Hb F variability in young children with Hb EE disease; and this can guide to appropriate care and counseling to affected families.
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Hemoglobinopatias , Talassemia beta , Humanos , Criança , Pré-Escolar , Estudos Retrospectivos , Polimorfismo de Nucleotídeo Único , Hemoglobina Fetal/genética , Hemoglobina Fetal/metabolismo , Hemoglobinopatias/genética , Mutação , Talassemia beta/genéticaRESUMO
Anemia is a major public health problem in many areas of Southeast Asia. Ascertaining anemia and defining its underlying causes is essential for providing appropriate care, management, and establishment of a control program. Limited studies on these have been carried out on people living at the borders of Thailand, Lao PDR, and Cambodia. This cross-sectional study was done in four areas along the borders of Thailand, Lao PDR, and Cambodia. Blood specimens were collected from subjects aged 15-18 years in four districts including Kantharalak, Si Sa Ket province (n = 36), Nam Khun (n = 109), Nam Yuen (n = 98), and Na Chaluai (n = 128), Ubon Ratchathani province, Thailand. RBC parameters were recorded, and serum ferritin (SF) level was measured. Diagnosis of thalassemia and hemoglobinopathies was based on hemoglobin (Hb) and DNA analyses. Measurement of C-reactive protein was performed to exclude false-negative result of iron deficiency. The prevalence of anemia was found to be 25.1%. ID accounted for only 10.5%. Various types of thalassemia were identified in 67.7% of the subjects. The overall prevalence of thalassemia included 3.5% α0-thalassemia, 0.8% ß-thalassemia, 47.7% Hb E, and 53.6% α+-thalassemia. The proportions of ID, thalassemia and combined ID and thalassemia among anemic subjects were 6.5%, 66.6%, and 20.4%, respectively. The results indicate that thalassemia and hemoglobinopathies rather than ID are major causes of anemia in Thailand-Lao PDR-Cambodia triangle. This information should prove useful for implementing an anemia control program in the regions.
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Anemia Ferropriva , Hemoglobinopatias , Deficiências de Ferro , Talassemia alfa , Talassemia beta , Humanos , Tailândia/epidemiologia , Estudos Transversais , Camboja/epidemiologia , Laos/epidemiologia , Hemoglobinopatias/genética , Talassemia alfa/complicações , Talassemia beta/complicaçõesRESUMO
OBJECTIVE: The degree of ineffective erythropoiesis is known to be associated with clinical severity among individuals with thalassemia. The association of ineffective erythropoiesis biomarker levels with different thalassemia genotypes, however, remains limited. The aim of this study was to explore the level of phosphatidylserine-exposed red blood cells (PS-exposed RBCs) and ineffective erythropoiesis biomarkers (growth-differentiation factor-15 and soluble transferrin receptors) in patients with different genotypes. METHODS: A cross-sectional study was conducted on 139 patients of age 18 years and above with different genotypes at Srinagarind Hospital, Khon Kaen University, Thailand. The levels of PS-exposed RBCs were determined using flow cytometry. Measurements of growth-differentiation factor-15 (GDF-15) and soluble transferrin receptors (sTfR) were evaluated by the ELISA method. RESULTS: The PS-exposed RBCs levels were found to be significantly higher in splenectomized beta-thalassemia patients. Patients with beta-thalassemia had the highest GDF-15 levels, followed by patients with non-deletional alpha-thalassemia. Patients with non-deletional alpha-thalassemia showed elevated hemoglobin levels and reduced GDF-15 levels after splenectomy. Patients with beta-thalassemia and non-deletional alpha-thalassemia had the highest levels of PS-exposed RBCs and ineffective erythropoiesis biomarkers, which correlated with the clinical severity of thalassemia. CONCLUSIONS: The levels of ineffective erythropoiesis biomarkers were different across thalassemia genotypes. Splenectomy may improve clinical symptoms of patients with non-deletional alpha thalassemia but not of patients with beta-thalassemia. These findings demonstrate differences in the degree of ineffective erythropoiesis in thalassemia, which emphasizes the need for different treatment approaches among patients with different thalassemia genotypes.
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OBJECTIVES: A co-inheritance of α0-thalassemia can ameliorate the clinical severity of the hemoglobin (Hb) E-ß-thalassemia disease. This information should be provided at prenatal diagnosis. Identification of α0-thalassemia in an affected fetus is therefore valuable. We have explored this genetic interaction in a large cohort of affected fetuses with hemoglobin (Hb) E-ß-thalassemia in northeast Thailand. METHODS: A study was done retrospectively on 1,592 couples at risk of having fetuses with Hb E-ß0-thalassemia, encountered from January 2011 to December 2019. A total of 415 left-over DNA specimens of the affected fetuses with Hb E-ß0-thalassemia disease were further investigated. Examination of α0-thalassemia was done using gap-PCR or a multiplex PCR assay for simultaneous detection of Hb E and α0-thalassemia mutations. RESULTS: Of the 415 affected fetuses, the two most common ß0-thalassemia genes found were the codons 41/42 (-TTCT) (199/415; 48.0%) and codon 17 (A-T) (115/415; 27.7%). α0-thalassemia was found unexpectedly in 21 (5.1%) fetuses. Hematologic phenotypes of the parents indicated that it was impossible to differentiate a pure ß0-thalassemia carrier from a double ß0-thalassemia/α0-thalassemia heterozygote unless DNA analysis is performed. In contrast, a reduced level of Hb E in the Hb E carrier (<25%) is a valuable marker for predicting double heterozygosity for Hb E/α0-thalassemia. This could be further confirmed using a multiplex PCR assay. CONCLUSIONS: There is a high prevalence of co-inheritance of α0-thalassemia in fetuses with Hb E-ß0-thalassemia disease. In a high-risk population such as Thailand, we recommend screening for α0-thalassemia in all affected fetuses with Hb E-ß0-thalassemia disease and providing complete genetic information to the parents to make appropriate decisions at prenatal diagnosis and genetic counseling.
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INTRODUCTION: Elevated hemoglobin (Hb) A2 is an important diagnostic marker for ß-thalassemia carriers. However, diagnosis of cases with borderline Hb A2 may be problematic. We described the molecular characteristics found in a large cohort of Thai subjects with borderline Hb A2. MATERIAL AND METHODS: Examination was done on 21,657 Thai subjects investigated for thalassemia at Khon Kaen University, Thailand. A total of 202 subjects with borderline Hb A2 (3.5-4.0%) were selectively recruited and hematological parameters were recorded. DNA variants in α-, ß-, δ-globin, and Krüppel-like factor 1 (KLF1) genes were examined using PCR. RESULTS: Among 202 subjects, DNA analysis identified carriers of α+-thalassemia (n = 48; 23.8%), ß-thalassemia (n = 22; 10.9%) and KLF1 mutations (n = 48; 23.8%). No molecular defect was observed in the remaining 84 (41.5%) subjects. Interaction of KLF1 and α-thalassemia was observed in 10 cases. Of the 22 ß-thalassemia carriers, five ß+-thalassemia mutations were identified with lower MCV and higher Hb A2. Seven KLF1 mutations were detected in 10 genotypes in subjects with higher MCV and Hb F. No ß0-thalassemia, α-globin gene triplication or δ-globin gene mutation was detected. CONCLUSIONS: A large proportion of subjects with borderline Hb A2 are not ß-thalassemia carriers and for those with ß-thalassemia, only mild ß+-thalassemia mutations were detected. Evaluation of the patients using Hb A2, Hb F and MCV values will help in selecting cases for further molecular analysis. The results should explain the unusual phenotype of the cases and facilitate a thalassemia screening program in the region.
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BACKGROUND: To evaluate whether the quantification of fetal hemoglobin (Hb) Bart's is useful for differentiation of α-thalassemia syndromes in the fetus and to characterize the fetal anemia associated with fetal α-hemoglobinopathy. METHODS: A total of 332 fetal blood specimens collected by cordocentesis were analyzed using capillary electrophoresis and the amount of Hb Bart's was recorded. The result was evaluated against thalassemia genotypes determined based on Hb and DNA analyses. Prenatal Hb and DNA characterization of the fetal anemia observed in two families was done. RESULTS: Among 332 fetuses investigated, Hb and DNA analyses identified 152 fetuses with normal genotypes. The remaining 180 fetuses carried α-thalassemia with several genotypes. Variable amounts of Hb Bart's were identified in all fetuses with α-thalassemia, which could be used for simple differentiation of fetal α-thalassemia genotypes. These included α+- and α0-thalassemia traits, homozygous α+-thalassemia and Hb Constant Spring (CS), Hb H disease, Hb H-CS and Hb H-Quong Sze diseases, homozygous α0-thalassemia causing the Hb Bart's hydrops fetalis and a remain uncharacterized α-thalassemia defect. The previously undescribed interactions of Hb Queens Park and Hb Amsterdam A1 with Hb E were detected in two fetuses with Hb Bart's of 0.5%. The Hb Queens Park-AEBart's disease was also noted in one pregnant woman. Prenatal analysis of the fetuses with severe fetal anemia and cardiomegaly with Hb Bart's of 9.0% and 13.6% revealed unexpectedly the homozygous Hb CS and a compound heterozygosity of Hb CS/Hb Pakse' with Hb E heterozygote, respectively. CONCLUSIONS: The usefulness of detecting and differentiation of fetal α-thalassemia syndromes by quantifying of Hb Bart's was demonstrated. Apart from the fatal condition of Hb Bart's hydrops fetalis associated with homozygous α0-thalassemia, homozygous Hb CS and a compound Hb CS/Hb Pakse' could result in severe fetal anemia and fetal complications, prenatal diagnosis is highly recommended. The simple Hb Bart's quantification of fetal blood should prove helpful in this matter.
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Talassemia alfa , Talassemia beta , Feminino , Feto , Hemoglobinas Anormais , Humanos , Hidropisia Fetal/diagnóstico , Hidropisia Fetal/genética , Gravidez , Diagnóstico Pré-Natal/métodos , Síndrome , Talassemia alfa/diagnóstico , Talassemia alfa/genéticaRESUMO
INTRODUCTION: Hemoglobinopathies are major public health problems worldwide. Accurate laboratory diagnosis of the carrier is essential, which includes initial screening, Hb analysis, and DNA analysis. For the first time, we have developed a single-tube quality control (QC) sample for these laboratory tests. METHODS: The QC sample was made from a lyophilized mixture of the stabilized hemolysate with carbon monoxide saturation and the white blood cells of known thalassemia mutations. Homogeneity and stability were examined by Hb and DNA analyses on day 0 and every month for 12 months, at room temperature, 4°C, and -20°C. A preliminary proficiency testing (PT) program for hemoglobinopathies using this single QC material was developed. RESULTS: Hemoglobin (Hb) and DNA analyses of a single-tube QC sample demonstrated satisfactory results of Hb analysis for at least five months and DNA analysis for at least one year of storage at -20°C. The results obtained from a preliminary PT program on five expert laboratories confirmed that a single tube QC sample prepared could be used as a PT item with various Hb and DNA analyses methods. CONCLUSION: A single lyophilized control sample has been generated for use in hemoglobinopathies' internal and external quality control program. Unlike other available control materials, which are used for individual testing, a single-tube QC sample generated can be used to control the pre-analytical and analytical processes of both Hb and DNA analyses and is suitable for use in the PT program of hemoglobinopathies.
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Hemoglobinopatias , Talassemia , DNA , Hemoglobinopatias/diagnóstico , Hemoglobinopatias/genética , Hemoglobinas/análise , Hemoglobinas/genética , Humanos , Controle de Qualidade , Talassemia/diagnósticoRESUMO
OBJECTIVE: Increased hemoglobin (Hb) A2 level is an important diagnostic marker for ß-thalassemia carrier screening. The level of Hb A2 is also useful for differentiating several thalassemia syndromes. We have examined data bases for reduced Hb A2 expression in a large cohort of Thai subjects. METHODS: A study was done on 1,498 subjects with non-thalassemia and various types of thalassemia and Hb variants to determine the effect of thalassemia genotypes and on 103 women of reproductive age to determine the effect of iron deficiency. Hb analysis was done using capillary electrophoresis, and thalassemia genotypes were defined by DNA analysis. Serum ferritin was measured using chemiluminescent microparticle immunoassay. RESULTS: Subjects were divided into 35 groups based on iron status, Hb, and DNA analysis. Decreased Hb A2 level was observed in those with Hb Q-Thailand, δ-hemoglobinopathies, δß0-thalassemia, Hb Lepore, iron deficiency, α-thalassemia, and especially Hb Constant Spring (Hb CS). While ß-thalassemia carriers with Hb H disease still had elevated Hb A2 levels, most of the ß-thalassemia carriers with Hb H-CS disease had Hb A2 less than 3.5% as a diagnostic cut-off. The lowest Hb A2 level was observed in those with Hb H-CS disease. CONCLUSION: Iron deficiency, Hb CS trait, homozygous Hb CS, and Hb H disease may reduce Hb A2 level, leading possibly to misdiagnosis of ß-thalassemia, especially in carriers with borderline Hb A2. Hb CS showed the strongest effect on Hb A2 expression. Understanding the basis for reduced Hb A2 expression may help reduce the diagnostic pitfalls of ß-thalassemia in the region.
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INTRODUCTION: Hemoglobin (Hb) analysis is a key testing for diagnosis of hemoglobinopathies. Accurate analysis, interpretation of results, and genetic risk assessment are important. We report on 8 years of the proficiency testing (PT) program for hemoglobinopathies in Thailand. METHODS: Laboratory participants were required to test two simulated PT items in each cycle using capillary electrophoresis, one was a husband and another was his pregnant wife. Related hematological parameters were provided. The participants also provide interpretation and evaluate the risk of having three severe thalassemia diseases in an expected fetus. Three cycles were operated per year in accordance with the ISO17043 and ISO13528 guidelines. A total of 84 laboratories throughout Thailand were participated. RESULTS: A total of 24 PT cycles were performed during 2012-2019. Most participants had Excellent performance for the PT items with normal, ß-thalassemia trait, hemoglobin E trait, hemoglobin E trait with α-thalassemia, and Hb H disease. However, when the PT items with homozygous Hb E and Hb E-ß-thalassemia were tested, an increase in a Needs improvement performance was noted. From 24 PT cycles, the performance with Excellent, Good, Fair, and Needs improvement was ranging from 10.5%-95.8%, 0%-11.3%, 0%-77.2%, and 2.3%-37.0%, respectively. CONCLUSION: Most participants have proven their performance to be reliable and demonstrated their abilities to provide interpretation and genetic risk assessment on most of the PT items. For complex thalassemia however, a need to improve the interpretation and risk assessment skills is required which is essential for effective prevention and control of severe thalassemia diseases in Thailand.
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Hemoglobinopatias/diagnóstico , Talassemia/diagnóstico , Eletroforese Capilar , Feminino , Testes Genéticos , Hemoglobina E/genética , Hemoglobinopatias/epidemiologia , Hemoglobinopatias/genética , Homozigoto , Humanos , Ensaio de Proficiência Laboratorial , Gravidez , Diagnóstico Pré-Natal , Tailândia/epidemiologia , Talassemia/epidemiologia , Talassemia/genéticaRESUMO
The δ-globin gene defects are clinically silent but interaction with ß-thalassemia can lead to a misdiagnosis of ß-thalassemia carrier. We report an extensive molecular characterization of δ-hemoglobinopathies in Thailand. Study was done on 32,108 subjects, encountered at the thalassemia screening. Six different approaches based on the reduced Hb A2 or appearance of Hb A2-derivative were established for selective recruitment of subjects. Among 32,108 subjects, a total of 296 subjects were suspected of having δ-globin gene defects. Of these 296 subjects, Hb and DNA analyses identified δ-hemoglobinopathies with 10 different mutations in 34 (0.11%) of them. These included a novel mutation, [δCD30(AGG>GGG) (n = 1)], 5 previously undescribed in Thailand, [δ-44(G>A) (n = 7), Hb A2-Troodos (n = 5), δIVSII-897(A>C) (n = 4), δ-68(C>T) (n = 2), and Hb A2-Indonesia (n = 1)], and 4 mutations previously found in Thailand, [Hb A2-Melbourne (n = 9), δ-77(T>C) (n = 3), Hb A2' (n = 1), and Hb A2-Kiriwong (n = 1)]. Genetic heterogeneities seen included interactions of δ-globin gene defects with heterozygous Hb E, ß-thalassemia, α-thalassemia, and in cis locations of the Hb A2-Troodos and Hb E mutations found for the first time. Rapid identification methods of these δ-globin gene mutations were developed. The results should prove useful to a prevention and control program of hemoglobinopathies in the region.
Assuntos
Talassemia/genética , Globinas delta/genética , Adolescente , Adulto , Criança , Feminino , Estudos de Associação Genética , Hemoglobina A2/genética , Humanos , Masculino , Mutação , Tailândia/epidemiologia , Talassemia/epidemiologia , Adulto JovemRESUMO
OBJECTIVES: Southeast Asian ovalocytosis (SAO) is an inherited red blood cell (RBC) membrane disorder, whereas hemoglobinopathies are inherited globin gene disorders. In an area where both diseases are prevalent, the interaction between them resulting in variable hematologic parameters can be encountered. However, little is known about the genetic interaction of SAO and thalassemia. We investigated the prevalence of SAO and hemoglobinopathy genotypes among newborns in southern Thailand. PATIENTS AND METHODS: This study was carried out on 297 newborns recruited consecutively at Naradhiwas Rajanagarindra Hospital in the south of Thailand. The SAO was identified on blood smear examination and polymerase chain reaction analysis. Thalassemia genotypes were defined. Hematologic parameters and hemoglobin (Hb) profiles were recorded and analyzed. RESULTS: Among 297 newborns, 15 (5.1%) carried SAO, whereas 70 (23.6%) had thalassemia with 15 different thalassemia genotypes. Abnormal Hb including Hb C, Hb Q-Thailand, and Hb D-Punjab were observed in 5 newborns. It was found in the nonthalassemic newborns that RBC count, Hb, and hematocrit of the nonthalassemic newborns with SAO were significantly lower than those without SAO. The same finding was also observed in the thalassemic newborns; RBC count, Hb, and hematocrit of the thalassemic newborns with SAO were significantly lower than those without SAO. However, the mean corpuscular volume, mean corpuscular Hb, and RBC distribution width of the SAO-newborns were significantly higher. CONCLUSIONS: Both SAO and hemoglobinopathy genotypes are common in southern Thailand. One should take this into consideration when evaluating neonatal anemia and other hematologic abnormalities. Identification of both genetic defects and long-term monitoring on the clinical outcome of this genetic interaction should be essential to understand the pathogenesis of these common genetic disorders in the region.
Assuntos
Eliptocitose Hereditária/sangue , Eliptocitose Hereditária/epidemiologia , Eliptocitose Hereditária/genética , Contagem de Eritrócitos , Hematócrito , Hemoglobina C/análise , Hemoglobina C/genética , Hemoglobinas Anormais/análise , Hemoglobinas Anormais/genética , Humanos , Recém-Nascido , Prevalência , Tailândia/epidemiologiaRESUMO
INTRODUCTION: A high frequency of ß-thalassemia in Lao People's Democratic Republic necessitates the importance of complete molecular data before a prevention and control program could be established. Limited data are available for Lao PDR. We have now reported an extended information on the molecular basis of ß-hemoglobinopathies in this population. METHODS: The study was done on 519 unrelated Laos subjects requested for thalassemia investigation. Hematological data were recorded. Hb profiles were obtained using a capillary electrophoresis system. α-And ß-globin genotyping was performed using PCR and related techniques. RESULTS: Among the 519 subjects, 287 (55.3%) were found to carry ß-hemoglobinopathies based on Hb and DNA analyses. These included Hb E carriers (n = 135), homozygous Hb E (n = 47), ß-thalassemia carriers (n = 70), Hb E-ß-thalassemia (n = 25), homozygous ß-thalassemia (n = 4), heterozygous δß0 -thalassemia (n = 2), and carriers of the ß-Hb variant (n = 3). Mutation analysis identified in addition to the Hb E, 8 different ß-thalassemia mutations including codon 17 (A-T), codons 41/42 (-TTCT), NT-28 (A-G), codons 71/72 (+A), IVS1-1 (G-T), 3.4 kb deletion, an initiation codon (T-G) and IVS2-654 (C-T). Two δß0 -thalassemia carriers (12.6 kb deletion) and three subjects with Hb Hope (ß136GGT-GAT ) were identified. Hematological features associated with these ß-hemoglobinopathies were presented. CONCLUSION: ß-hemoglobinopathies in the Laos population is heterogeneous. This information is relevant for setting up a molecular diagnostics and can provide a basis for genetic counseling and enable prenatal diagnosis.