The Evolving Role of Next-Generation Sequencing in Screening and Diagnosis of Hemoglobinopathies

Achour, Ahlem; Koopmann, Tamara T.; Baas, Frank; Harteveld, Cornelis L.

doi:10.3389/fphys.2021.686689

Cited by 27 publications

(12 citation statements)

References 33 publications

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“…Now with the evolving technology of Next Generation Sequencing many molecular diagnostic labs have implemented Inherited Disease Panels, Whole Exome‐ and Whole Genome Sequencing 28 for a variety of genetic diseases. For the hemoglobinopathies the high homology between the HBA1 and HBA2 and between HBD and HBB , puts a challenge to NGS methods as short sequence reads interfere with the specificity of the highly homologous genes.…”

Section: Molecular Diagnosticsmentioning

confidence: 99%

“…This may eventually be replaced by Next Generation Sequencing (NGS) based Copy Number Variation (CNV) detection, which becomes increasingly successful using better software tools to interpret NGS obtained data. 27 Now with the evolving technology of Next Generation Sequencing many molecular diagnostic labs have implemented Inherited Disease Panels, Whole Exome-and Whole Genome Sequencing 28 for a variety of genetic diseases. For the hemoglobinopathies the high homology between the HBA1 and HBA2 and between HBD and HBB, puts a challenge to NGS methods as short sequence reads interfere with the specificity of the highly homologous genes.…”

Section: Molecular Diagnosticsmentioning

confidence: 99%

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The hemoglobinopathies, molecular disease mechanisms and diagnostics

Harteveld

Achour

Arkesteijn

et al. 2022

Int J Lab Hematology

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Hemoglobinopathies are the most common monogenic disorders in the world with an ever increasing global disease burden each year. As most hemoglobinopathies show recessive inheritance carriers are usually clinically silent. Programmes for preconception and antenatal carrier screening, with the option of prenatal diagnosis are considered beneficial in many endemic countries. With the development of genetic tools such as Array analysis and Next Generation Sequencing in addition to state of the art screening at the hematologic, biochemic and genetic level, have contributed to the discovery of an increasing number of rare rearrangements and novel factors influencing the disease severity over the recent years. This review summarizes the basic requirements for adequate carrier screening analysis, the importance of genotype-phenotype correlation and how this may lead to the unrevealing exceptional interactions causing a clinically more severe phenotype in otherwise asymptomatic carriers. A special group of patients are β-thalassemia carriers presenting with features of β-thalassemia intermedia of various clinical severity. The disease mechanisms may involve duplicated α-globin genes, mosaic partial Uniparental Isodisomy of chromosome 11p15.4 where the HBB gene is located or haploinsufficiency of a non-linked gene SUPT5H on chromosome 19q, first described in two Dutch families with β-thalassemia trait without variants in the HBB gene.

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Section: Molecular Diagnosticsmentioning

confidence: 99%

Section: Molecular Diagnosticsmentioning

confidence: 99%

The hemoglobinopathies, molecular disease mechanisms and diagnostics

Harteveld

Achour

Arkesteijn

et al. 2022

Int J Lab Hematology

Self Cite

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“…Compared to its counterparts, such as MLPA and Comparative Genomic Hybridization (CGH) analysis, NGS has been reported to be less accurate in these cases. Therefore, best practice recommends that NGS to always be paired with Gap-PCR to compensate for this shortcoming, and Sanger sequencing to remain a mandatory confirmatory test ( 64 ). Further, the advent of long molecule third-generation sequencing technology may also offer a similar solution by directly reading the entire length of the gene sequence with no apparent GC preference ( 65 , 66 ).…”

Section: Discussionmentioning

confidence: 99%

Applications of next generation sequencing in the screening and diagnosis of thalassemia: A mini-review

et al. 2022

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Thalassemias are a group of inherited blood disorders that affects 5–7% of the world population. Comprehensive screening strategies are essential for the management and prevention of this disorder. Today, many clinical and research laboratories have widely utilized next-generation sequencing (NGS) technologies to identify diseases, from germline and somatic disorders to infectious diseases. Yet, NGS application in thalassemia is limited and has just recently surfaced due to current demands in seeking alternative DNA screening tools that are more efficient, versatile, and cost-effective. This review aims to understand the several aspects of NGS technology, including its most current and expanding uses, advantages, and limitations, along with the issues and solutions related to its integration into routine screening and diagnosis of thalassemias. Hitherto, NGS has been a groundbreaking technology that offers tremendous improvements as a diagnostic tool for thalassemia in terms of its higher throughput, accuracy, and adaptability. The superiority of NGS in detecting rare variants, solving complex hematological problems, and providing non-invasive alternatives to neonatal diagnosis cannot be overlooked. However, several pitfalls still preclude its use as a stand-alone technique over conventional methods.

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“…However, these techniques are still time consuming and are currently considered more costly because sequencing for thalassemia involves few genes. 25 Although rapid advances in genetic techniques enable simpler and more comprehensive diagnostics in thalassemia, we believe that the evaluation of hematological features and hemoglobin pattern analysis will still remain fundamental for accurate and resource efficient sample handling. In addition, the understanding of hematological abnormalities and results of hemoglobin pattern analysis relative to genotype is important to avoid misinterpretation of genetic results.…”

Section: Discussionmentioning

confidence: 99%

Additional value of red blood cell parameters in predicting uncommon α‐thalassemia; experience from 10 years of α‐globin gene sequencing and copy number variation analysis

Fjeld

Sudmann-Day

Grimholt

et al. 2022

Int J Lab Hematology

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Introduction The diagnosis of rare forms of α‐thalassemia requires laborious genetic analyses. Accurate sample selection for such evaluation is therefore essential. The main objectives of this study were to investigate the predictive power of red blood cell parameters to detect rare forms of α‐thalassemia (substudy 1), and to explore the frequency of rare versus common forms of α‐thalassemia in our sample population (substudy 2). Methods In substudy 1, we reviewed all blood samples selected for extended α‐hemoglobinopathy evaluation at our laboratory during 2011–2020 (n = 1217), which included DNA sequencing and/or copy number variation analysis. We assessed α‐thalassemia positive samples at different levels of mean corpuscular hemoglobin (MCH) alone and in combination with results for red blood cell count (RBC) or red cell distribution width (RDW). In substudy 2, we examined the distribution of α‐thalassemia genotypes for all samples submitted to a first‐tier hemoglobinopathy evaluation at our laboratory during 2014–2020 (n = 6495). Results In substudy 1, both RBC and RDW added predictive value in detecting rare forms of α‐thalassemia in samples from adults and children. In adult samples with MCH ≤ 23 pg, the presence of erythrocytosis increased the detection rate from 27% to 74% as compared to non‐erythrocytosis, while normal RDW increased the detection rate from 36% to 86% as compared to elevated RDW. In substudy 2, rare forms of α‐thalassemia were detected in 12% of α‐thalassemia positive samples. Conclusion Initial assessment of MCH, RBC, and RDW provided valuable predictive information about the presence of rare forms of α‐thalassemia during hemoglobinopathy evaluation.

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The Evolving Role of Next-Generation Sequencing in Screening and Diagnosis of Hemoglobinopathies

Cited by 27 publications

References 33 publications

The hemoglobinopathies, molecular disease mechanisms and diagnostics

The hemoglobinopathies, molecular disease mechanisms and diagnostics

Applications of next generation sequencing in the screening and diagnosis of thalassemia: A mini-review

Additional value of red blood cell parameters in predicting uncommon α‐thalassemia; experience from 10 years of α‐globin gene sequencing and copy number variation analysis

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