Molecular diagnosis of thrombocytopenia-absent radius syndrome using next-generation sequencing.
BACKGROUND
Thrombocytopenia-absent radius (TAR) syndrome is a rare autosomal recessive disease. Patients are compound heterozygotes for a loss-of-function allele, which in most cases is a large genomic deletion on chromosome 1q21.1 containing the RBM8A gene, and a noncoding variant located in the 5'UTR (rs139428292) or intronic (rs201779890) regions of RBM8A. As the molecular genetic testing in TAR requires multiple techniques for detection of copy-number variations (CNV) and nucleotide substitutions, we tested whether a next-generation sequencing (NGS) approach could identify both alterations.


METHODS
Two unrelated families were analyzed with Ion PGM sequencing using a target panel of genes responsible for different forms of inherited thrombocytopenia. A statistical quantitative evaluation of amplicon coverage was performed to detect CNV, in particular those on the RBM8A gene.


RESULTS
All the probands were apparently homozygous for the rare allele inherited by the father at the rs139428292 locus, suggesting the presence of a deletion on the maternal chromosome. The statistical analysis confirmed the hemizygous condition of RBM8A.


CONCLUSIONS
We concluded that NGS approaches could be used as a cost-effective method for molecular investigation of TAR as they could simultaneously detect CNV and point mutations.
Department of Medical Sciences, University of Trieste, Trieste, Italy.
E
E
Nicchia
University of Bari, Bari, Italy.
P
P
Giordano
Institute for Maternal and Child Health - IRCCS Burlo Garofolo, Trieste, Italy.
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C
Greco
Institute for Maternal and Child Health - IRCCS Burlo Garofolo, Trieste, Italy.
D
D
De Rocco
Department of Medical Sciences, University of Trieste, Trieste, Italy.|Institute for Maternal and Child Health - IRCCS Burlo Garofolo, Trieste, Italy.
A
A
Savoia
D000328:Adult; D002872:Chromosome Deletion; D000080984:Congenital Bone Marrow Failure Syndromes; D056915:DNA Copy Number Variations / Q000235:genetics; D005190:Family; D005260:Female; D006801:Humans; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods / Q000592:standards*; D016601:RNA-Binding Proteins / Q000235:genetics*; D011884:Radius; D017422:Sequence Analysis, DNA / Q000379:methods / Q000592:standards; D013921:Thrombocytopenia / Q000175:diagnosis* / Q000235:genetics; D038062:Upper Extremity Deformities, Congenital / Q000175:diagnosis* / Q000235:genetics
C406243:RBM8A protein, human; D016601:RNA-Binding Proteins
Thrombocytopenia-absent radius (TAR) syndrome; copy-number variations (CNVs); ion PGM sequencing; next-generation sequencing (NGS); point mutation