Identification of Duchenne/Becker muscular dystrophy mosaic carriers through a combined DNA/RNA analysis

Stefania Zampatti, Julia Mela, Cristina Peconi, Giulia Pagliaroli, Stefania Carboni, Giuseppe Barrano, Ilaria Zito, Raffaella Cascella, Gianluca Marella, Filippo Milano, Mauro Arcangeli, Carlo Caltagirone, Antonio Novelli, Emiliano Giardina

Research output: Contribution to journalArticle

Abstract

OBJECTIVE: The Duchenne/Becker muscular dystrophy carrier screening includes the evaluation of mutations in DMD gene and the most widely used analysis is the MLPA for the DMD deletions/duplications detection. The high frequency of de novo mutations permits to estimate a risk up to 20% of mosaicisms for mothers of sporadic DMD children. The purpose of this study is to evaluate alternative analytical strategy for the detection of mosaics carrier women, in order to improve the recurrence risk estimation.

METHOD: Different DNA and RNA analyses were conducted on samples from a woman that conceived a DMD fetus without previous family history of dystrophynopathy.

RESULTS: Standard MLPA analysis failed to identify mosaicism, even if MLPA doses suggested it. Electrophoresis and direct sequencing conducted on RNA permitted to detect two different amplicons of cDNAs, demonstrating the presence of somatic mosaicism. Subsequent detection of a second affected fetus confirmed the mosaic status on the mother.

CONCLUSION: The implementation of RNA analysis in diagnostic algorithm can increase the sensitivity of carrier test for mothers of sporadic affected patients, permitting detection of mosaic status. A revision of analytical guidelines is needed in order to improve the recurrence risk estimation and support prenatal genetic counseling.

Original languageEnglish
JournalPrenatal Diagnosis
DOIs
Publication statusE-pub ahead of print - Oct 10 2018

Fingerprint

Mosaicism
Duchenne Muscular Dystrophy
Mothers
RNA
DNA
Fetus
Recurrence
Mutation
Genetic Counseling
Electrophoresis
Complementary DNA
Guidelines
Genes

Cite this

Identification of Duchenne/Becker muscular dystrophy mosaic carriers through a combined DNA/RNA analysis. / Zampatti, Stefania; Mela, Julia; Peconi, Cristina; Pagliaroli, Giulia; Carboni, Stefania; Barrano, Giuseppe; Zito, Ilaria; Cascella, Raffaella; Marella, Gianluca; Milano, Filippo; Arcangeli, Mauro; Caltagirone, Carlo; Novelli, Antonio; Giardina, Emiliano.

In: Prenatal Diagnosis, 10.10.2018.

Research output: Contribution to journalArticle

Zampatti, S, Mela, J, Peconi, C, Pagliaroli, G, Carboni, S, Barrano, G, Zito, I, Cascella, R, Marella, G, Milano, F, Arcangeli, M, Caltagirone, C, Novelli, A & Giardina, E 2018, 'Identification of Duchenne/Becker muscular dystrophy mosaic carriers through a combined DNA/RNA analysis', Prenatal Diagnosis. https://doi.org/10.1002/pd.5369
Zampatti, Stefania ; Mela, Julia ; Peconi, Cristina ; Pagliaroli, Giulia ; Carboni, Stefania ; Barrano, Giuseppe ; Zito, Ilaria ; Cascella, Raffaella ; Marella, Gianluca ; Milano, Filippo ; Arcangeli, Mauro ; Caltagirone, Carlo ; Novelli, Antonio ; Giardina, Emiliano. / Identification of Duchenne/Becker muscular dystrophy mosaic carriers through a combined DNA/RNA analysis. In: Prenatal Diagnosis. 2018.
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abstract = "OBJECTIVE: The Duchenne/Becker muscular dystrophy carrier screening includes the evaluation of mutations in DMD gene and the most widely used analysis is the MLPA for the DMD deletions/duplications detection. The high frequency of de novo mutations permits to estimate a risk up to 20{\%} of mosaicisms for mothers of sporadic DMD children. The purpose of this study is to evaluate alternative analytical strategy for the detection of mosaics carrier women, in order to improve the recurrence risk estimation.METHOD: Different DNA and RNA analyses were conducted on samples from a woman that conceived a DMD fetus without previous family history of dystrophynopathy.RESULTS: Standard MLPA analysis failed to identify mosaicism, even if MLPA doses suggested it. Electrophoresis and direct sequencing conducted on RNA permitted to detect two different amplicons of cDNAs, demonstrating the presence of somatic mosaicism. Subsequent detection of a second affected fetus confirmed the mosaic status on the mother.CONCLUSION: The implementation of RNA analysis in diagnostic algorithm can increase the sensitivity of carrier test for mothers of sporadic affected patients, permitting detection of mosaic status. A revision of analytical guidelines is needed in order to improve the recurrence risk estimation and support prenatal genetic counseling.",
author = "Stefania Zampatti and Julia Mela and Cristina Peconi and Giulia Pagliaroli and Stefania Carboni and Giuseppe Barrano and Ilaria Zito and Raffaella Cascella and Gianluca Marella and Filippo Milano and Mauro Arcangeli and Carlo Caltagirone and Antonio Novelli and Emiliano Giardina",
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T1 - Identification of Duchenne/Becker muscular dystrophy mosaic carriers through a combined DNA/RNA analysis

AU - Zampatti, Stefania

AU - Mela, Julia

AU - Peconi, Cristina

AU - Pagliaroli, Giulia

AU - Carboni, Stefania

AU - Barrano, Giuseppe

AU - Zito, Ilaria

AU - Cascella, Raffaella

AU - Marella, Gianluca

AU - Milano, Filippo

AU - Arcangeli, Mauro

AU - Caltagirone, Carlo

AU - Novelli, Antonio

AU - Giardina, Emiliano

N1 - This article is protected by copyright. All rights reserved.

PY - 2018/10/10

Y1 - 2018/10/10

N2 - OBJECTIVE: The Duchenne/Becker muscular dystrophy carrier screening includes the evaluation of mutations in DMD gene and the most widely used analysis is the MLPA for the DMD deletions/duplications detection. The high frequency of de novo mutations permits to estimate a risk up to 20% of mosaicisms for mothers of sporadic DMD children. The purpose of this study is to evaluate alternative analytical strategy for the detection of mosaics carrier women, in order to improve the recurrence risk estimation.METHOD: Different DNA and RNA analyses were conducted on samples from a woman that conceived a DMD fetus without previous family history of dystrophynopathy.RESULTS: Standard MLPA analysis failed to identify mosaicism, even if MLPA doses suggested it. Electrophoresis and direct sequencing conducted on RNA permitted to detect two different amplicons of cDNAs, demonstrating the presence of somatic mosaicism. Subsequent detection of a second affected fetus confirmed the mosaic status on the mother.CONCLUSION: The implementation of RNA analysis in diagnostic algorithm can increase the sensitivity of carrier test for mothers of sporadic affected patients, permitting detection of mosaic status. A revision of analytical guidelines is needed in order to improve the recurrence risk estimation and support prenatal genetic counseling.

AB - OBJECTIVE: The Duchenne/Becker muscular dystrophy carrier screening includes the evaluation of mutations in DMD gene and the most widely used analysis is the MLPA for the DMD deletions/duplications detection. The high frequency of de novo mutations permits to estimate a risk up to 20% of mosaicisms for mothers of sporadic DMD children. The purpose of this study is to evaluate alternative analytical strategy for the detection of mosaics carrier women, in order to improve the recurrence risk estimation.METHOD: Different DNA and RNA analyses were conducted on samples from a woman that conceived a DMD fetus without previous family history of dystrophynopathy.RESULTS: Standard MLPA analysis failed to identify mosaicism, even if MLPA doses suggested it. Electrophoresis and direct sequencing conducted on RNA permitted to detect two different amplicons of cDNAs, demonstrating the presence of somatic mosaicism. Subsequent detection of a second affected fetus confirmed the mosaic status on the mother.CONCLUSION: The implementation of RNA analysis in diagnostic algorithm can increase the sensitivity of carrier test for mothers of sporadic affected patients, permitting detection of mosaic status. A revision of analytical guidelines is needed in order to improve the recurrence risk estimation and support prenatal genetic counseling.

U2 - 10.1002/pd.5369

DO - 10.1002/pd.5369

M3 - Article

JO - Prenatal Diagnosis

JF - Prenatal Diagnosis

SN - 0197-3851

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