MLPA followed by target-NGS to detect mutations in the dystrophin gene of Peruvian patients suspected of DMD/DMB

María Luisa Guevara-Fujita; Francia Huaman-Dianderas; Daisy Obispo; Rodrigo Sánchez; Victor Barrenechea; Diana Rojas-Málaga; Alejandro Estrada-Cuzcano; Milana Trubnykova; Mario Cornejo-Olivas; Victoria Marca; Bertha Gallardo; Milagros Dueñas-Roque; Ana Protzel; Carlos Castañeda; Hugo Abarca; Luis Celis; Jorge La Serna-Infantes; Ricardo Fujita

doi:10.1002/mgg3.1759

MLPA followed by target-NGS to detect mutations in the dystrophin gene of Peruvian patients suspected of DMD/DMB

María Luisa Guevara-Fujita, Francia Huaman-Dianderas, Daisy Obispo, Rodrigo Sánchez, Victor Barrenechea, Diana Rojas-Málaga, Alejandro Estrada-Cuzcano, Milana Trubnykova, Mario Cornejo-Olivas, Victoria Marca, Bertha Gallardo, Milagros Dueñas-Roque, Ana Protzel, Carlos Castañeda, Hugo Abarca, Luis Celis, Jorge La Serna-Infantes, Ricardo Fujita

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Abstract

Background: We report the molecular analysis of the DMD gene in a group of Peruvian patients with Duchenne/Becker dystrophinopathy. This is the first study to thoroughly characterize mutations in this population. Methods: We used the combination of multiplex ligation-dependent probe amplification (MLPA) and sequencing analysis of the DMD gene. We recruited Peruvian patients in 2 years from reference national hospitals. We performed DNA tests in 152 patients, checking first exon deletion/duplication by MLPA, and subsequently, if negative, samples were sequenced to detect point mutations. Results: The average age for diagnosis was 9.8 years, suggesting a delay for timely diagnosis and care. We found causal DMD mutations in 125 patients: 72 (57.6%) exon deletions/duplications (41.6% deletions, 16.0% duplications), and 53 (42.4%) point mutations (27.2% nonsense, 9.6% small indels, and 5.6% splice site). Conclusion: Due to our genetic background, we expected a higher number of novel and recurrent causal mutations in our sample. Results showed 16% of novel mutations, similar to other well-studied populations.

Original language	English
Article number	e1759
Journal	Molecular genetics & genomic medicine
Volume	9
Issue number	9
DOIs	https://doi.org/10.1002/mgg3.1759
State	Published - Sep 2021

Bibliographical note

Funding Information:
The authors thank the participants and their relatives, the ADM-Peru, the physicians, including R. Yábar, J. Toro, A. Tori, R. Caparó, S. Samalvides, G. Chávez, and M. Chávez, and others who helped with sample collection and clinical exams. They also thank PTC Therapeutics and authorities at the Facultad de Medicina, Universidad de San Martín de Porres, who helped fund this study.

Publisher Copyright:
© 2021 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals LLC

Keywords

Becker muscular dystrophy
Duchenne muscular dystrophy
molecular diagnosis
multiple ligation probe amplification
targeted Next Generation Sequencing

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Guevara-Fujita, M. L., Huaman-Dianderas, F., Obispo, D., Sánchez, R., Barrenechea, V., Rojas-Málaga, D., Estrada-Cuzcano, A., Trubnykova, M., Cornejo-Olivas, M., Marca, V., Gallardo, B., Dueñas-Roque, M., Protzel, A., Castañeda, C., Abarca, H., Celis, L., La Serna-Infantes, J., & Fujita, R. (2021). MLPA followed by target-NGS to detect mutations in the dystrophin gene of Peruvian patients suspected of DMD/DMB. Molecular genetics & genomic medicine, 9(9), [e1759]. https://doi.org/10.1002/mgg3.1759

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title = "MLPA followed by target-NGS to detect mutations in the dystrophin gene of Peruvian patients suspected of DMD/DMB",

abstract = "Background: We report the molecular analysis of the DMD gene in a group of Peruvian patients with Duchenne/Becker dystrophinopathy. This is the first study to thoroughly characterize mutations in this population. Methods: We used the combination of multiplex ligation-dependent probe amplification (MLPA) and sequencing analysis of the DMD gene. We recruited Peruvian patients in 2 years from reference national hospitals. We performed DNA tests in 152 patients, checking first exon deletion/duplication by MLPA, and subsequently, if negative, samples were sequenced to detect point mutations. Results: The average age for diagnosis was 9.8 years, suggesting a delay for timely diagnosis and care. We found causal DMD mutations in 125 patients: 72 (57.6%) exon deletions/duplications (41.6% deletions, 16.0% duplications), and 53 (42.4%) point mutations (27.2% nonsense, 9.6% small indels, and 5.6% splice site). Conclusion: Due to our genetic background, we expected a higher number of novel and recurrent causal mutations in our sample. Results showed 16% of novel mutations, similar to other well-studied populations.",

keywords = "Becker muscular dystrophy, Duchenne muscular dystrophy, molecular diagnosis, multiple ligation probe amplification, targeted Next Generation Sequencing",

author = "Guevara-Fujita, {Mar{\'i}a Luisa} and Francia Huaman-Dianderas and Daisy Obispo and Rodrigo S{\'a}nchez and Victor Barrenechea and Diana Rojas-M{\'a}laga and Alejandro Estrada-Cuzcano and Milana Trubnykova and Mario Cornejo-Olivas and Victoria Marca and Bertha Gallardo and Milagros Due{\~n}as-Roque and Ana Protzel and Carlos Casta{\~n}eda and Hugo Abarca and Luis Celis and {La Serna-Infantes}, Jorge and Ricardo Fujita",

note = "Publisher Copyright: {\textcopyright} 2021 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals LLC",

year = "2021",

month = sep,

doi = "10.1002/mgg3.1759",

language = "Ingl{\'e}s",

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Guevara-Fujita, ML, Huaman-Dianderas, F, Obispo, D, Sánchez, R, Barrenechea, V, Rojas-Málaga, D, Estrada-Cuzcano, A, Trubnykova, M, Cornejo-Olivas, M, Marca, V, Gallardo, B, Dueñas-Roque, M, Protzel, A, Castañeda, C, Abarca, H, Celis, L, La Serna-Infantes, J & Fujita, R 2021, 'MLPA followed by target-NGS to detect mutations in the dystrophin gene of Peruvian patients suspected of DMD/DMB', Molecular genetics & genomic medicine, vol. 9, no. 9, e1759. https://doi.org/10.1002/mgg3.1759

TY - JOUR

T1 - MLPA followed by target-NGS to detect mutations in the dystrophin gene of Peruvian patients suspected of DMD/DMB

AU - Guevara-Fujita, María Luisa

AU - Huaman-Dianderas, Francia

AU - Obispo, Daisy

AU - Sánchez, Rodrigo

AU - Barrenechea, Victor

AU - Rojas-Málaga, Diana

AU - Estrada-Cuzcano, Alejandro

AU - Trubnykova, Milana

AU - Cornejo-Olivas, Mario

AU - Marca, Victoria

AU - Gallardo, Bertha

AU - Dueñas-Roque, Milagros

AU - Protzel, Ana

AU - Castañeda, Carlos

AU - Abarca, Hugo

AU - Celis, Luis

AU - La Serna-Infantes, Jorge

AU - Fujita, Ricardo

PY - 2021/9

Y1 - 2021/9

N2 - Background: We report the molecular analysis of the DMD gene in a group of Peruvian patients with Duchenne/Becker dystrophinopathy. This is the first study to thoroughly characterize mutations in this population. Methods: We used the combination of multiplex ligation-dependent probe amplification (MLPA) and sequencing analysis of the DMD gene. We recruited Peruvian patients in 2 years from reference national hospitals. We performed DNA tests in 152 patients, checking first exon deletion/duplication by MLPA, and subsequently, if negative, samples were sequenced to detect point mutations. Results: The average age for diagnosis was 9.8 years, suggesting a delay for timely diagnosis and care. We found causal DMD mutations in 125 patients: 72 (57.6%) exon deletions/duplications (41.6% deletions, 16.0% duplications), and 53 (42.4%) point mutations (27.2% nonsense, 9.6% small indels, and 5.6% splice site). Conclusion: Due to our genetic background, we expected a higher number of novel and recurrent causal mutations in our sample. Results showed 16% of novel mutations, similar to other well-studied populations.

AB - Background: We report the molecular analysis of the DMD gene in a group of Peruvian patients with Duchenne/Becker dystrophinopathy. This is the first study to thoroughly characterize mutations in this population. Methods: We used the combination of multiplex ligation-dependent probe amplification (MLPA) and sequencing analysis of the DMD gene. We recruited Peruvian patients in 2 years from reference national hospitals. We performed DNA tests in 152 patients, checking first exon deletion/duplication by MLPA, and subsequently, if negative, samples were sequenced to detect point mutations. Results: The average age for diagnosis was 9.8 years, suggesting a delay for timely diagnosis and care. We found causal DMD mutations in 125 patients: 72 (57.6%) exon deletions/duplications (41.6% deletions, 16.0% duplications), and 53 (42.4%) point mutations (27.2% nonsense, 9.6% small indels, and 5.6% splice site). Conclusion: Due to our genetic background, we expected a higher number of novel and recurrent causal mutations in our sample. Results showed 16% of novel mutations, similar to other well-studied populations.

KW - Becker muscular dystrophy

KW - Duchenne muscular dystrophy

KW - molecular diagnosis

KW - multiple ligation probe amplification

KW - targeted Next Generation Sequencing

UR - http://www.scopus.com/inward/record.url?scp=85111526488&partnerID=8YFLogxK

U2 - 10.1002/mgg3.1759

DO - 10.1002/mgg3.1759

M3 - Artículo

AN - SCOPUS:85111526488

SN - 2324-9269

VL - 9

JO - Molecular genetics & genomic medicine

JF - Molecular genetics & genomic medicine

IS - 9

M1 - e1759

ER -

MLPA followed by target-NGS to detect mutations in the dystrophin gene of Peruvian patients suspected of DMD/DMB

Abstract

Bibliographical note

Keywords

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