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Identification of Duchenne/Becker muscular dystrophy mosaic carriers through a combined DNA/RNA analysis
Author(s) -
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
Publication year - 2018
Publication title -
prenatal diagnosis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.956
H-Index - 97
eISSN - 1097-0223
pISSN - 0197-3851
DOI - 10.1002/pd.5369
Subject(s) - multiplex ligation dependent probe amplification , duchenne muscular dystrophy , prenatal diagnosis , multiplex , amplicon , germline mosaicism , biology , genetics , muscular dystrophy , multiplex polymerase chain reaction , medicine , mutation , polymerase chain reaction , fetus , gene , pregnancy , exon
Abstract Objective The Duchenne/Becker muscular dystrophy (DMD) carrier screening includes the evaluation of mutations in DMD gene, and the most widely used analysis is the multiplex ligation‐dependent probe amplification (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.