Application of Targeted Next-Generation Sequencing for the Investigation of Thalassemia in a Developing Country: A Single Center Experience

Abstract

Thalassemia is identified as a prevalent disease in Malaysia, known to be one of the developing countries. Fourteen patients with confirmed cases of thalassemia were recruited from the Hematology Laboratory. The molecular genotypes of these patients were tested using the multiplex-ARMS and GAP-PCR methods. The samples were repeatedly investigated using the Devyser Thalassemia kit (Devyser, Sweden), a targeted NGS panel targeting the coding regions of hemoglobin genes, namely the HBA1, HBA2, and HBB genes, which were used in this study. There were many different genetic variants found in 14 unrelated cases. Out of all fourteen cases, NGS was able to determine an additional -50 G>A (HBB:c.-100G>A) that were not identified by the multiplex-ARMS method, including HBA2 mutations, namely CD 79 (HBA2:c.239C>G). Other than that, CD 142 (HBA2:c.427T>C) and another non-deletional alpha thalassemia and alpha triplication were also not picked up by the GAP-PCR methods. We illustrated a broad, targeted NGS-based test that proposes benefits rather than using traditional screening or basic molecular methods. The results of this study should be heeded, as this is the first report on the practicality of targeted NGS concerning the biological and phenotypic features of thalassemia, especially in a developing population. Discovering rare pathogenic thalassemia variants and additional secondary modifiers may facilitate precise diagnosis and better disease prevention.

Current Status of Newborn Bloodspot Screening Worldwide 2024: A Comprehensive Review of Recent Activities (2020–2023)

“…Researchers have taken advantage of MS/MS technology to review and develop reference ranges for Carnitine-Acylcarnitine Translocase (CACT) deficiency and Carnitine Palmitoyl Transferase 2 (CPT II) deficiency [ 585 ]. Other research has centered on experiences in applying NGS to screening for thalassemia [ 586 ], validating a POC screening test for G6PDD [ 587 ], the need for additional training when setting up a new condition based on experiences with CH in 12 government hospitals and 20 health clinics [ 588 ], and the need for a mandated IEM-related course for certain healthcare disciplines at the university level [ 589 ].…”

Unlocking Insights into Alloantibodies in Thalassemia: Findings from a Single-Center Study

“…As per current estimates, around 6.8% of Malaysians are thalassemia carriers, which puts them at risk of developing anemia to varying degrees 1 . Hb E thalassemia and beta thalassemia are the predominant inherited hematological disorders of beta-globin in Malaysia 2, 3 . Patients with severe thalassemia rely on blood transfusions every 3-4 weeks to treat anemia 4 .According to the current guidelines of the Thalassemia International Federation, blood transfusions are recommended for pre-transfusion Hb levels of 9-10 g/dL in patients with transfusion-dependent thalassemia 5 .…”

Characterization of New Alpha Zero (α0) Thalassaemia Deletion (−−GB) among Malays in Malaysian Population

“…Alpha (α)-thalassaemia is the most common inherited disorder resulting from a reduced rate of synthesis of the α-globin chain. The prevalence of alpha thalassaemia trait in local studies including the Southeast Asia regions is higher (4.08% to 48.9%) [1][2][3][4][5][6] compared to beta (β)-thalassaemia carriers, which are reported by many researchers and range from 0.93% to 12.8% [1,2,5,7]. In one meta-analysis study involving 83,864 thousand subjects in the Southeast Asian region, the prevalence of α-thalassaemia in Malaysia is 17.4%, which is lower compared to the overall prevalence in the Southeast Asian region, which is 22.6% [8].…”