Development and validation of a comprehensive mutation and deletion detection assay for SDHB, SDHC, and SDHD.
BACKGROUND
Lack of sequencing validation and complexity of deletion testing hinder genetic diagnosis of SDH-associated paraganglioma/pheochromocytoma.


METHODS
We developed sequencing assays and multiplex ligation-dependent probe amplification (MLPA) deletion detection for SDHB, SDHC and SDHD. Clinical performance was validated on 141 blinded samples, previously tested at NIH.


RESULTS
Sequencing and deletion detection were highly reproducible and agreed with previous NIH results in 99.3% and 100%, respectively.


CONCLUSIONS
DNA sequencing combined with MLPA allows reliable and simplified genotyping of SDHB, SDHC and SDHD.
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN 55902, USA.
Dragana
D
Milosevic
Patrick
P
Lundquist
Kendall
K
Cradic
Noemi
N
Vidal-Folch
ThanhTruc
T
Huynh
Karel
K
Pacak
Stefan K G
SK
Grebe
D000310:Adrenal Gland Neoplasms / Q000175:diagnosis / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D008565:Membrane Proteins / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D010235:Paraganglioma / Q000235:genetics; D010673:Pheochromocytoma / Q000175:diagnosis / Q000235:genetics; D016133:Polymerase Chain Reaction; D017384:Sequence Deletion / Q000235:genetics; D013385:Succinate Dehydrogenase / Q000235:genetics*
D008565:Membrane Proteins; C108096:SDHC protein, human; C491545:SDHD protein, human; C108095:SDHB protein, human; D013385:Succinate Dehydrogenase