Accurate Molecular Diagnosis of Gaucher Disease Using Clinical Exome Sequencing as a First-Tier Test

Accurate Molecular Diagnosis of Gaucher Disease Using Clinical Exome Sequencing as a First-Tier Test

Gaucher disease (GD) is an autosomal recessive lysosomal disorder due to beta-glucosidase gene (<i>GBA</i>) mutations. The molecular diagnosis of GD is complicated by the presence of recombinant alleles originating from a highly homologous pseudogene. Clinical exome sequencing (CES) is a...

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Main Authors: Stefania Zampieri, Silvia Cattarossi, Eleonora Pavan, Antonio Barbato, Agata Fiumara, Paolo Peruzzo, Maurizio Scarpa, Giovanni Ciana, Andrea Dardis
Format: Article
Language:English
Published: MDPI AG 2021-05-01
Series:International Journal of Molecular Sciences
Subjects:
<i>GBA</i>
clinical exome sequencing
MLPA
Online Access:https://www.mdpi.com/1422-0067/22/11/5538
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spelling doaj-023a1100b64a4c90ae3116e53dadc8862021-06-01T00:57:40ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-05-01225538553810.3390/ijms22115538Accurate Molecular Diagnosis of Gaucher Disease Using Clinical Exome Sequencing as a First-Tier TestStefania Zampieri0Silvia Cattarossi1Eleonora Pavan2Antonio Barbato3Agata Fiumara4Paolo Peruzzo5Maurizio Scarpa6Giovanni Ciana7Andrea Dardis8Regional Coordinator Centre for Rare Diseases, University Hospital of Udine, 33100 Udine, ItalyRegional Coordinator Centre for Rare Diseases, University Hospital of Udine, 33100 Udine, ItalyRegional Coordinator Centre for Rare Diseases, University Hospital of Udine, 33100 Udine, ItalyDepartment of Clinical Medicine and Surgery, Federico II University Hospital, 80131 Naples, ItalyPediatric Unit, Regional Referral Center for Inherited Metabolic Disease, University of Catania, 95123 Catania, ItalyRegional Coordinator Centre for Rare Diseases, University Hospital of Udine, 33100 Udine, ItalyRegional Coordinator Centre for Rare Diseases, University Hospital of Udine, 33100 Udine, ItalyRegional Coordinator Centre for Rare Diseases, University Hospital of Udine, 33100 Udine, ItalyRegional Coordinator Centre for Rare Diseases, University Hospital of Udine, 33100 Udine, ItalyGaucher disease (GD) is an autosomal recessive lysosomal disorder due to beta-glucosidase gene (<i>GBA</i>) mutations. The molecular diagnosis of GD is complicated by the presence of recombinant alleles originating from a highly homologous pseudogene. Clinical exome sequencing (CES) is a rapid genetic approach for identifying disease-causing mutations. However, copy number variation and recombination events are poorly detected, and further investigations are required to avoid mis-genotyping. The aim of this work was to set-up an integrated strategy for GD patients genotyping using CES as a first-line test. Eight patients diagnosed with GD were analyzed by CES. Five patients were fully genotyped, while three were revealed to be homozygous for mutations that were not confirmed in the parents. Therefore, MLPA (multiplex ligation-dependent probe amplification) and specific long-range PCR were performed, and two recombinant alleles, one of them novel, and one large deletion were identified. Furthermore, an MLPA assay performed in one family resulted in the identification of an additional novel mutation (p.M124V) in a relative, in trans with the known p.N409S mutation. In conclusion, even though CES has become extensively used in clinical practice, our study emphasizes the importance of a comprehensive molecular strategy to provide proper <i>GBA</i> genotyping and genetic counseling.https://www.mdpi.com/1422-0067/22/11/5538<i>GBA</i>clinical exome sequencingMLPA
collection DOAJ
language English
format Article
sources DOAJ
author Stefania Zampieri
Silvia Cattarossi
Eleonora Pavan
Antonio Barbato
Agata Fiumara
Paolo Peruzzo
Maurizio Scarpa
Giovanni Ciana
Andrea Dardis
spellingShingle Stefania Zampieri
Silvia Cattarossi
Eleonora Pavan
Antonio Barbato
Agata Fiumara
Paolo Peruzzo
Maurizio Scarpa
Giovanni Ciana
Andrea Dardis
Accurate Molecular Diagnosis of Gaucher Disease Using Clinical Exome Sequencing as a First-Tier Test
International Journal of Molecular Sciences
<i>GBA</i>
clinical exome sequencing
MLPA
author_facet Stefania Zampieri
Silvia Cattarossi
Eleonora Pavan
Antonio Barbato
Agata Fiumara
Paolo Peruzzo
Maurizio Scarpa
Giovanni Ciana
Andrea Dardis
author_sort Stefania Zampieri
title Accurate Molecular Diagnosis of Gaucher Disease Using Clinical Exome Sequencing as a First-Tier Test
title_short Accurate Molecular Diagnosis of Gaucher Disease Using Clinical Exome Sequencing as a First-Tier Test
title_full Accurate Molecular Diagnosis of Gaucher Disease Using Clinical Exome Sequencing as a First-Tier Test
title_fullStr Accurate Molecular Diagnosis of Gaucher Disease Using Clinical Exome Sequencing as a First-Tier Test
title_full_unstemmed Accurate Molecular Diagnosis of Gaucher Disease Using Clinical Exome Sequencing as a First-Tier Test
title_sort accurate molecular diagnosis of gaucher disease using clinical exome sequencing as a first-tier test
publisher MDPI AG
series International Journal of Molecular Sciences
issn 1661-6596
1422-0067
publishDate 2021-05-01
description Gaucher disease (GD) is an autosomal recessive lysosomal disorder due to beta-glucosidase gene (<i>GBA</i>) mutations. The molecular diagnosis of GD is complicated by the presence of recombinant alleles originating from a highly homologous pseudogene. Clinical exome sequencing (CES) is a rapid genetic approach for identifying disease-causing mutations. However, copy number variation and recombination events are poorly detected, and further investigations are required to avoid mis-genotyping. The aim of this work was to set-up an integrated strategy for GD patients genotyping using CES as a first-line test. Eight patients diagnosed with GD were analyzed by CES. Five patients were fully genotyped, while three were revealed to be homozygous for mutations that were not confirmed in the parents. Therefore, MLPA (multiplex ligation-dependent probe amplification) and specific long-range PCR were performed, and two recombinant alleles, one of them novel, and one large deletion were identified. Furthermore, an MLPA assay performed in one family resulted in the identification of an additional novel mutation (p.M124V) in a relative, in trans with the known p.N409S mutation. In conclusion, even though CES has become extensively used in clinical practice, our study emphasizes the importance of a comprehensive molecular strategy to provide proper <i>GBA</i> genotyping and genetic counseling.
topic <i>GBA</i>
clinical exome sequencing
MLPA
url https://www.mdpi.com/1422-0067/22/11/5538
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