ONDRISeq VCF files of the ONDRI cases were imported into VarSeq® (Golden Helix, Bozeman, MT, United States) and variants were annotated with sequence ontologies. Minor allele frequencies (MAFs) were obtained from the Genome Aggregation Database (gnomAD v.2.0.1v3 non-neuro)37 (link). Rare (MAF < 0.01), nonsynonymous variants were prioritized. Further assessment of variants was performed to identify those in genes known to contribute to Mendelian forms of the patient’s disease of diagnosis and those classified as pathogenic or likely pathogenic in ClinVar38 (link), Online Mendelian Inheritance in Man (OMIM)39 , and/or the Alzforum Mutation Database40 . All identified variants were considered those likely to be contributing to Mendelian forms of disease.
All samples were genotyped for C9orf72 using both amplicon length analysis and repeat-primed polymerase chain reaction (PCR), as previously described41 (link). Harbouring >30 repeats is a commonly accepted genetic cause of ALS and FTD41 (link),42 (link), and therefore was the cutoff used to determine those with pathogenic repeat expansions.
All samples were genotyped for C9orf72 using both amplicon length analysis and repeat-primed polymerase chain reaction (PCR), as previously described41 (link). Harbouring >30 repeats is a commonly accepted genetic cause of ALS and FTD41 (link),42 (link), and therefore was the cutoff used to determine those with pathogenic repeat expansions.
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