Rapid detection and prenatal diagnosis of β-thalassaemia: studies in Indian and Cypriot populations in the UK

Old, John; Varawalla, Nermeen; Weatherall, D. J.

doi:10.1016/0140-6736(90)92338-i

Cited by 283 publications

(145 citation statements)

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“…Genomic DNA was isolated from whole blood in anticoagulant (EDTA) by using the standard proteinase K-phenol-chloroform procedure as described in Old et al [8]. Abnormal hemoglobins were analyzed by agarose gel electrophoresis at pH 8.6, Hb A 2 and Hb F were determined by gel elution and alkali denaturation [9], respectively.…”

Section: Phenotype Analysismentioning

confidence: 99%

Spectrum of β‐thalassemia mutations and their association with allelic sequence polymorphisms at the β‐globin gene cluster in an eastern Indian population

et al. 2002

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In this report, the spectrum of ␤-thalassemia mutations and genotype-to-phenotype correlations were defined in large number of patients (␤-thalassemia carriers and major) with varying disease severity in an Eastern Indian population mainly from the state of West Bengal. The five most common ␤-thalassemia mutations were detected, which included IVS1-5 (G➝C), codon 15 (G➝A), codon 26 (G➝A), codon 30 (G➝C), and codon 41/42 (−TCTT). These accounted for 85% in 80 ␤-thalassemic alleles deciphered from 56 patients, including ␤-thalassemia major and carriers, and 15% of alleles remained uncharacterized in these patients. Expression of the human ␤-globin gene is regulated by an array of cis-acting DNA elements, including five DNase I hypersensitive sites (HSs) in the locus control region (LCR), promoters that incorporate certain silencer elements, and enhancers at 3 of the ␤-globin gene. For detailed studies and to understand the molecular basis of ␤-thalassemia, we studied two groups of subjects: a group of 12 patients from four families having ␤-thalassemia major and carrier phenotype and a control group of 26 healthy individuals. In these two groups, we examined portions of the ␤-globin gene locus control region HSs 1, 2, 3, and 4, which included the (CA) x (TA) y repeat motif, the (AT) x N y (AT) z repeat motif, the inverted repeat sequence TGGGGACCCCA, the promoter region of the G ␥-globin gene, an (AT) x (T) y repeat 5 of the silencer region, and the ␤-globin gene and its 3 flanking region. We investigated the allelic sequence polymorphisms in these regions and their association with the ␤-thalassemia mutations to know the possible genotype-phenotype relationship in ␤-thalassemia patients. An analysis of cisacting regulatory regions showed varied sequence haplotypes associated with some frequent ␤-thalassemia mutations in this Eastern Indian population. Am.

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Section: Phenotype Analysismentioning

confidence: 99%

Spectrum of β‐thalassemia mutations and their association with allelic sequence polymorphisms at the β‐globin gene cluster in an eastern Indian population

et al. 2002

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“…Genomic DNA was isolated from whole blood in anticoagulant (EDTA) by using proteinase KÀphe-nolÀchloroform standard procedure [3]. Hemoglobin variants were analysed by agarose gel electrophoresis at pH 8.6; HbA 2 was determined by gel elution and HbF by alkali denaturation, respectively [4].…”

Section: Phenotype Analysismentioning

confidence: 99%

“…ARMS-PCR method was used to detect known 23 b-thalassemia mutations [3,5]. b-globin gene cluster locus control 5¢ HS 2, 3, and 4 ($460 bp, $446 bp, and $442 bp from GeneBank coordinates 8757À 9217, 4397À4991, and 668À1109) were sequenced by the ABI Prism 377 automated DNA sequencer using dye terminators chemistry.…”

Section: Mutation Analysis and Examination Of B-lcr Hssmentioning

confidence: 99%

Study of the single nucleotide polymorphism (SNP) at the palindromic sequence of hypersensitive site (HS)4 of the human β‐globin locus control region (LCR) in Indian population

et al. 2001

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LCR, a genetic regulatory element, was examined in b-thalassemia patients who do not show any mutation in the b-globin genes. We sequenced LCR-HS2, HS3, and HS4 in samples from 16 such patients from the Indian population and found only one SNP A-G in the inverted repeat in HS4. A signi®cant association was observed between the G allele and occurrence of b-thalassemia by Fisher's exact test. The AG and GG genotypes showed higher relative risk as compared to the AA genotype. We also observed linkage disequilibrium between the A/G polymorphism and the AT-rich motif of the LCR HS2 region, suggesting that the G allele could be an evolutionarily new mutation in the study population. Am.

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“…The ARMS-PCR technique is a popular technique to identify the known b-thalassemia mutations [16,17]. In this study, we have successfully devised a multiplex ARMS-PCR system for the detection of b-thalassemia mutations commonly found in AfricanAmericans and Asians, including those of Chinese, Taiwanese, Southeast Asian, and Asian Indian descent.…”

Section: Discussionmentioning

confidence: 99%

Molecular genetic confirmatory testing from newborn screening samples for the common African‐American, Asian Indian, Southeast Asian, and Chinese β‐thalassemia mutations

et al. 2005

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b-Thalassemia is a serious health problem in the United States, especially in California, due to increased Asian immigration. Neonatal screening by using high-performance liquid chromatography (HPLC) or isoelectric focusing (IEF) may lead to confusion due to interactions of various hemoglobinopathies with b-thalassemia. Our purpose was to develop single-tube multiplexed PCR assays using original neonatal screening specimens to identify the mutations responsible for b-thalassemia in order to expedite diagnostic confirmation. Primers were designed for two to six common ethnic-specific mutations using the amplification refractory mutation system (ARMS). This multiplex ARMS approach was standardized using DNA samples with known mutations for b-thalassemia in those of Asian (Southeast Asian, Chinese, and Asian Indian) and African-American descent. Specimens from African-American neonates were tested for two mutations (À88 and À29); Asian Indians for five mutations (IVSI-1, IVSI-5, codons (Cd) 41/42, Cd 8/9, and 619-bp deletion); Chinese, Taiwanese, and Southeast Asians for seven mutations (Cd 41/42, Cd 17, À28, IVSII-654, Cd 71/72, IVSI-5, and IVSI-1). We identified each of these b-thalassemia mutations in multiplexed ARMS from positive control samples. We tested 25 anonymized dried blood specimens from neonates who had been diagnosed with b-thalassemia and who also belonged to these ethnic groups. We detected a mutation specific to the neonate's ethnic group using the ARMS approach in nearly all specimens, and the results were confirmed by sequencing. Multiplexed ARMS for ethnic-specific b-thalassemia mutations from the original newborn screening dried blood specimens is a rapid and efficient approach for diagnostic confirmation. Am.

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Rapid detection and prenatal diagnosis of β-thalassaemia: studies in Indian and Cypriot populations in the UK

Cited by 283 publications

References 10 publications

Spectrum of β‐thalassemia mutations and their association with allelic sequence polymorphisms at the β‐globin gene cluster in an eastern Indian population

Spectrum of β‐thalassemia mutations and their association with allelic sequence polymorphisms at the β‐globin gene cluster in an eastern Indian population

Study of the single nucleotide polymorphism (SNP) at the palindromic sequence of hypersensitive site (HS)4 of the human β‐globin locus control region (LCR) in Indian population

Molecular genetic confirmatory testing from newborn screening samples for the common African‐American, Asian Indian, Southeast Asian, and Chinese β‐thalassemia mutations

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