Study of pathophysiology and molecular characterization of congenital anemia in India using targeted next-generation sequencing approach

Int J Hematol. 2019 Nov;110(5):618-626. doi: 10.1007/s12185-019-02716-9. Epub 2019 Aug 10.


Most patients with anemia are diagnosed through clinical phenotype and basic laboratory testing. Nonetheless, in cases of rare congenital anemias, some patients remain undiagnosed despite undergoing an exhaustive workup. Genetic testing is complicated by the large number of genes that are involved in rare anemias, due to similarities in the clinical presentation. We sought to enhance the diagnosis of patients with congenital anemias by using targeted next-generation sequencing. The genetic diagnosis was performed by gene capture followed by next-generation sequencing of 76 genes known to cause anemia syndromes. Genetic diagnosis was achieved in 17 of 21 transfusion-dependent patients and undiagnosed by conventional workup. Four cases were diagnosed with red cell membrane protein defects, four patients were diagnosed with pyruvate kinase deficiency, one case of adenylate kinase deficiency, one case of glucose phosphate isomerase deficiency, one case of hereditary xerocytosis, three cases having combined membrane and enzyme defect, two cases with Diamond-Blackfan anemia (DBA) and 1 with CDA type II with 26 different mutations, of which 21 are novel. Earlier incorporation of this NGS method into the workup of patients with congenital anemia may improve patient care and enable genetic counselling.

Keywords: Congenital anemia; Erythroenzymopathies; India; Membranopathies; Molecular genetics; Next-generation sequencing.

MeSH terms

  • Adenylate Kinase / genetics
  • Anemia / congenital*
  • Anemia / genetics
  • Anemia, Diamond-Blackfan / genetics
  • Anemia, Hemolytic, Congenital / genetics
  • Anemia, Hemolytic, Congenital Nonspherocytic / genetics
  • Cytokines / genetics
  • Genetic Testing / methods*
  • Glucose-6-Phosphate Isomerase / genetics
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Hydrops Fetalis / genetics
  • India
  • Mutation*
  • Pyruvate Kinase / deficiency
  • Pyruvate Kinase / genetics
  • Pyruvate Metabolism, Inborn Errors / genetics


  • Cytokines
  • Pyruvate Kinase
  • Adenylate Kinase
  • GPI protein, human
  • Glucose-6-Phosphate Isomerase

Supplementary concepts

  • Pyruvate Kinase Deficiency of Red Cells
  • Xerocytosis, hereditary