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Updated: Jun 12, 2026
Rare Event Detection Using Error-corrected DNA and RNA Sequencing
Published on: August 3, 2018
DNA sequencing errors in molecular diagnostics of filamin myopathy
Zagaa Odgerel1, Peter F M van der Ven, Dieter O Fürst
1National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892-9404, USA.
Background:
Filamin myopathy is a neuromuscular disorder manifesting with predominantly limb-girdle muscle weakness and in many patients with diaphragm paralysis and cardiomyopathy, caused by mutations in the filamin C (FLNC) gene. Molecular diagnosis of filamin myopathy based on direct DNA sequencing of coding exons is compromised by the presence of a high homology pseudogene (pseFLNC) located approximately 53.6 kb downstream of the functional FLNC gene on chromosome 7q.
Methods:
Molecular cloning, RT-PCR and real-time PCR methods were used to detect sequence differences between the FLNC and pseFLNC that are implicated in known or potential molecular diagnostic errors. Overall, 50 patients with a phenotype resembling filamin myopathy have been screened for mutations in FLNC.
Results:
FLNC sequence inconsistencies caused by the interference from pseFLNC were identified and diagnostic errors involving, in particular, the detection of the most frequent disease-causing FLNC p.W2710X mutation resolved. Mismatches between the FLNC and pseFLNC sequences were tabulated for future use.
Conclusions:
We devise a strategy that allows one to discern mutations occurring in the functional FLNC from those harbored in pseFLNC, thus preventing possible complications in future research and patient genetic testing.
Insights
Filamin myopathy diagnosis is improved by a new strategy distinguishing mutations in the FLNC gene from its pseudogene (pseFLNC). This prevents errors in genetic testing for this neuromuscular disorder.
Area of Science:
- Genetics
- Molecular Biology
- Neuromuscular Disorders
Background:
- Filamin myopathy, caused by FLNC gene mutations, presents with muscle weakness, diaphragm paralysis, and cardiomyopathy.
- Accurate molecular diagnosis is hindered by a highly homologous FLNC pseudogene (pseFLNC).
Purpose of the Study:
- To develop a method to differentiate between the functional FLNC gene and its pseudogene (pseFLNC).
- To resolve diagnostic errors in filamin myopathy genetic testing caused by pseudogene interference.
Main Methods:
- Utilized molecular cloning, RT-PCR, and real-time PCR to identify sequence differences between FLNC and pseFLNC.
- Screened 50 patients with filamin myopathy phenotypes for FLNC mutations.
Main Results:
- Identified FLNC sequence inconsistencies due to pseFLNC interference, resolving diagnostic errors.
- Successfully distinguished disease-causing FLNC mutations from those in pseFLNC, including the common p.W2710X mutation.
- Tabulated sequence mismatches between FLNC and pseFLNC for future diagnostic applications.
Conclusions:
- Devised a strategy to accurately discern mutations in the functional FLNC gene from its pseudogene.
- Prevented complications in future research and patient genetic testing for filamin myopathy.
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