Next-generation sequencing unravels homozygous mutation in glucose-6-phosphate isomerase, GPI c.1040G &gt; A (p.Arg347His) causing hemolysis in an Indian infant

Jamwal, Manu; Aggarwal, Anuradha; Das, Anirban; Maitra, Arindam; Sharma, Prashant; Krishnan, Shekhar; Arora, Neeraj; Bansal, Deepak; Das, Reena

doi:10.1016/j.cca.2017.02.012

Cited by 20 publications

(18 citation statements)

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“…Additionally, the usefulness of Sanger sequencing is limited for the diagnoses of complex, multi-gene disorders or those with locus heterogeneity. Recent advances in molecular technologies have helped to identify unexpected candidate genes in numerous inherited disorders including IHA [438990]. Various NGS-based methods have been developed, including whole genome sequencing, exome sequencing, and gene panels [91].…”

Section: Next-generation Sequencing For the Genetic Diagnosis Of Ihamentioning

confidence: 99%

Diagnostic approaches for inherited hemolytic anemia in the genetic era

2017

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Inherited hemolytic anemias (IHAs) are genetic diseases that present with anemia due to the increased destruction of circulating abnormal RBCs. The RBC abnormalities are classified into the three major disorders of membranopathies, hemoglobinopathies, and enzymopathies. Traditional diagnosis of IHA has been performed via a step-wise process combining clinical and laboratory findings. Nowadays, the etiology of IHA accounts for germline mutations of the responsible genes coding for the structural components of RBCs. Recent advances in molecular technologies, including next-generation sequencing, inspire us to apply these technologies as a first-line approach for the identification of potential mutations and to determine the novel causative genes in patients with IHAs. We herein review the concept and strategy for the genetic diagnosis of IHAs and provide an overview of the preparations for clinical applications of the new molecular technologies.

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Section: Next-generation Sequencing For the Genetic Diagnosis Of Ihamentioning

confidence: 99%

Diagnostic approaches for inherited hemolytic anemia in the genetic era

2017

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“…The G6PD gene, located in the long arm of chromosome X (Xq28), consists of 13 exons and 12 introns encoding 515 amino acids. Over 200 G6PD mutations have been reported worldwide, and the distribution of mutations is of racial and regional heterogeneity 11 , 12 . Up to now, the epidemiological data of neonatal G6PD deficiency in China remains elusive 13 , 14 .…”

Section: Introductionmentioning

confidence: 99%

Newborn screening of glucose-6-phosphate dehydrogenase deficiency in Guangxi, China: determination of optimal cutoff value to identify heterozygous female neonates

Luo

Li³

et al. 2018

Sci Rep

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The aim of this study is to assess the disease incidence and mutation spectrum of glucose-6-phosphate dehydrogenase (G6PD) deficiency in Guangxi, China, and to determine an optimal cutoff value to identify heterozygous female neonates. A total of 130, 635 neonates were screened from the year of 2013 to 2017. Neonates suspected for G6PD deficiency were further analyzed by quantitatively enzymatic assay and G6PD mutation analysis. The overall incidence of G6PD deficiency was 7.28%. A total of 14 G6PD mutations were identified, and different mutations lead to varying levels of G6PD enzymatic activities. The best cut-off value of G6PD activity in male subjects is 2.2 U/g Hb, same as conventional setting. In female population, however, the cut-off value is found to be 2.8 U/g Hb (sensitivity: 97.5%, specificity: 87.7%, AUC: 0.964) to best discriminate between normal and heterozygotes, and 1.6 U/g Hb (sensitivity: 82.2%, specificity: 85.9%, AUC: 0.871) between heterozygotes and deficient subjects. In conclusion, we have conducted a comprehensive newborn screening of G6PD deficiency in a large cohort of population from Guangxi, China, and first established a reliable cut-off value of G6PD activity to distinguish heterozygous females from either normal or deficient subjects.

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“…Until now, only three patients of haemolytic anaemia associated with hyperbilirubinaemia and splenomegaly due to GPI deficiency were reported from India. Molecular diagnosis in the cases of North India with HNSHA associated with neurological deficit showed c.1040G>A (p.Arg347His) was identified by NGS 3. We have previously reported one novel mutation c.1459C>T (p.Leu487Phe) mutation from our laboratory in exon 16 by Sanger sequencing 14.…”

Section: Discussionmentioning

confidence: 97%

Molecular diagnosis of unexplained haemolytic anaemia using targeted next-generation sequencing panel revealed (p.Ala337Thr) novel mutation in GPI gene in two Indian patients

et al. 2018

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Glucose-6-phosphate isomerase (GPI) deficiency is an autosomal recessive genetic disorder causing congenital haemolytic anaemia (CHA). Diagnosis of GPI deficiency by the biochemical method is unpredicted. Molecular diagnosis by identifying genetic mutation is the gold standard method for confirmation of disease, but causative genes involved in CHA are numerous, and identifying a gene-by-gene approach using Sanger sequencing is also cumbersome, expensive and labour intensive. Recently, next-generation targeted sequencing is more useful in the diagnosis of unexplained haemolytic anaemia. We used targeted next-generation sequencing (NGS) clinical panel for diagnosis of unexplained haemolytic anaemia in two Indian patients which were pending for a long time. All possible causes of haemolytic anaemia were found within normal limit. NGS by clinical exome panel revealed homozygous novel missense mutation in exon 12, c.1009G>A (p.Ala337Thr) in both patients. We further confirm by measuring red blood cell GPI activity in the patients and showed deficiency whereas parents were having intermediate activity. c.1009G>A mutation was also confirmed by Sanger sequencing of exon 12 of GPI gene. The structural–functional analysis by bioinformatics software like Swiss PDB, PolyPhen-2 and PyMol suggested that this pathogenic variant has a direct impact on the structural rearrangement at the region near the active site of the enzyme. This rapid and high-performance targeted NGS assay can be configured to detect specific CHA mutations unique to an individual defect, making it a potentially valuable method for diagnosis of unexplained haemolytic anaemia.

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Next-generation sequencing unravels homozygous mutation in glucose-6-phosphate isomerase, GPI c.1040G > A (p.Arg347His) causing hemolysis in an Indian infant

Cited by 20 publications

References 10 publications

Diagnostic approaches for inherited hemolytic anemia in the genetic era

Diagnostic approaches for inherited hemolytic anemia in the genetic era

Newborn screening of glucose-6-phosphate dehydrogenase deficiency in Guangxi, China: determination of optimal cutoff value to identify heterozygous female neonates

Molecular diagnosis of unexplained haemolytic anaemia using targeted next-generation sequencing panel revealed (p.Ala337Thr) novel mutation in GPI gene in two Indian patients

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