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Array comparative genomic hybridization identifies a heterozygous deletion of exon 3 of the RYR2 gene.

Leong IU, Sucich J, Prosser DO, Skinner JR, Crawford JR, Higgins C, Love DR - Ups. J. Med. Sci. (2015)

Bottom Line: The standard diagnostic screening involves Sanger-based sequencing of 45 of the 105 translated exons of the RYR2 gene, and copy number changes of a limited number of exons that are detected using multiplex ligation-dependent probe amplification (MLPA).The CGH array detected a 1.1 kb deletion encompassing exon 3 of the RYR2 gene.The aCGH method offers significant advantages over MLPA in genetic screening for heritable cardiac disorders.

View Article: PubMed Central - PubMed

Affiliation: Diagnostic Genetics, LabPLUS, Auckland City Hospital , PO Box 110031, Auckland 1142 , New Zealand.

ABSTRACT

Background: Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a heritable cardiac disorder characterized by life-threatening ventricular tachycardia caused by exercise or acute emotional stress. The standard diagnostic screening involves Sanger-based sequencing of 45 of the 105 translated exons of the RYR2 gene, and copy number changes of a limited number of exons that are detected using multiplex ligation-dependent probe amplification (MLPA).

Methods: In the current study, a previously validated bespoke array comparative genomic hybridization (aCGH) technique was used to detect copy number changes in the RYR2 gene in a 43-year-old woman clinically diagnosed with CPVT.

Results: The CGH array detected a 1.1 kb deletion encompassing exon 3 of the RYR2 gene. This is the first report using the aCGH technique to screen for mutations causing CPVT.

Conclusions: The aCGH method offers significant advantages over MLPA in genetic screening for heritable cardiac disorders.

No MeSH data available.


Related in: MedlinePlus

Copy number changes in chromosome 1q43 of the proband. DEVA software output showing a copy number change (deletion; 10 probes; log2ratio: –0.6726) localized to chromosome 1q43 (235,560,602-235,561,416; hg18 co-ordinates) for the proband (II-3).
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Figure 3: Copy number changes in chromosome 1q43 of the proband. DEVA software output showing a copy number change (deletion; 10 probes; log2ratio: –0.6726) localized to chromosome 1q43 (235,560,602-235,561,416; hg18 co-ordinates) for the proband (II-3).

Mentions: The aCGH analysis of the proband identified a heterozygous deletion on chromosome 1 (1q43) that was approximately 816 bp in size (hg18 co-ordinates chr1:235,560,602-235,561,416) (Figure 3). This deletion encompasses the whole of exon 3 of the RYR2 gene with flanking intronic regions (Figure 4). Sanger-based sequencing was used to confirm the aCGH results and also to determine the exact breakpoints of the deletion. Primers were designed to flank the region of interest (Figure 4A), and subsequent Sanger-based sequencing of the resulting smaller amplicon confirmed the aCGH results but with greater resolution such that the deletion proved to be 1,126 bp in size (Figure 4A). This mutation (c.169-198_c.273+823 del1126) has been previously reported by Ohno et al. (12) and Szentpali et al. (Figure 5) (13). Exon 3 of the RYR2 gene encodes for a highly conserved region of the RYR2 protein, and the mutation is an in-frame deletion of 35 amino acids (p.Asn57-Gly91).


Array comparative genomic hybridization identifies a heterozygous deletion of exon 3 of the RYR2 gene.

Leong IU, Sucich J, Prosser DO, Skinner JR, Crawford JR, Higgins C, Love DR - Ups. J. Med. Sci. (2015)

Copy number changes in chromosome 1q43 of the proband. DEVA software output showing a copy number change (deletion; 10 probes; log2ratio: –0.6726) localized to chromosome 1q43 (235,560,602-235,561,416; hg18 co-ordinates) for the proband (II-3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526874&req=5

Figure 3: Copy number changes in chromosome 1q43 of the proband. DEVA software output showing a copy number change (deletion; 10 probes; log2ratio: –0.6726) localized to chromosome 1q43 (235,560,602-235,561,416; hg18 co-ordinates) for the proband (II-3).
Mentions: The aCGH analysis of the proband identified a heterozygous deletion on chromosome 1 (1q43) that was approximately 816 bp in size (hg18 co-ordinates chr1:235,560,602-235,561,416) (Figure 3). This deletion encompasses the whole of exon 3 of the RYR2 gene with flanking intronic regions (Figure 4). Sanger-based sequencing was used to confirm the aCGH results and also to determine the exact breakpoints of the deletion. Primers were designed to flank the region of interest (Figure 4A), and subsequent Sanger-based sequencing of the resulting smaller amplicon confirmed the aCGH results but with greater resolution such that the deletion proved to be 1,126 bp in size (Figure 4A). This mutation (c.169-198_c.273+823 del1126) has been previously reported by Ohno et al. (12) and Szentpali et al. (Figure 5) (13). Exon 3 of the RYR2 gene encodes for a highly conserved region of the RYR2 protein, and the mutation is an in-frame deletion of 35 amino acids (p.Asn57-Gly91).

Bottom Line: The standard diagnostic screening involves Sanger-based sequencing of 45 of the 105 translated exons of the RYR2 gene, and copy number changes of a limited number of exons that are detected using multiplex ligation-dependent probe amplification (MLPA).The CGH array detected a 1.1 kb deletion encompassing exon 3 of the RYR2 gene.The aCGH method offers significant advantages over MLPA in genetic screening for heritable cardiac disorders.

View Article: PubMed Central - PubMed

Affiliation: Diagnostic Genetics, LabPLUS, Auckland City Hospital , PO Box 110031, Auckland 1142 , New Zealand.

ABSTRACT

Background: Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a heritable cardiac disorder characterized by life-threatening ventricular tachycardia caused by exercise or acute emotional stress. The standard diagnostic screening involves Sanger-based sequencing of 45 of the 105 translated exons of the RYR2 gene, and copy number changes of a limited number of exons that are detected using multiplex ligation-dependent probe amplification (MLPA).

Methods: In the current study, a previously validated bespoke array comparative genomic hybridization (aCGH) technique was used to detect copy number changes in the RYR2 gene in a 43-year-old woman clinically diagnosed with CPVT.

Results: The CGH array detected a 1.1 kb deletion encompassing exon 3 of the RYR2 gene. This is the first report using the aCGH technique to screen for mutations causing CPVT.

Conclusions: The aCGH method offers significant advantages over MLPA in genetic screening for heritable cardiac disorders.

No MeSH data available.


Related in: MedlinePlus