The NM_003494.4: c.5057+5G>A variant in DYSF, which is also known as NM_001130987.2: c.5174+5G>A, occurs within the splice donor motif of intron 45. RNAseq and cDNA analysis has demonstrated that this variant disrupts splicing, resulting in a frameshift and premature truncation, with no detectable normally spliced transcript (PMID: 36983702, 10766988; PVS1_RNA). This variant has been identified in at least two patients with LGMD, including in a homozygous state (0.25 pts, PMID: 10766988) and in unknown phase with a pathogenic variant (c.5979dup p.(Glu1994ArgfsTer3), 0.5 pts, PMID: 36983702) (PM3_Supporting). At least one patient with this variant displayed progressive limb girdle muscle weakness and severely reduced or absent dysferlin protein expression, which is highly specific for DYSF-associated LGMD (PP4_Moderate, PMID: 36983702, 10766988). The variant was shown to co-segregate with the LGMD phenotype in five affected members of a consanguineous family of Yemenite Jewish descent (PP1_Moderate; PMID: 10766988). The highest population minor allele frequency is 0.00002196 (2/91068 exome alleles) in the South Asian population in gnomAD v4.1.0, which is less than the LGMD VCEP threshold (≤0.0001) for PM2_Supporting, meeting this criterion (PM2_Supporting). In summary, this variant meets the criteria to be classified as Pathogenic for autosomal recessive limb girdle muscular dystrophy based on the ACMG/AMP criteria applied, as specified by the ClinGen LGMD VCEP (LGMD VCEP specifications version 1.0.0; 03/18/2025): PVS1_RNA, PM3_Supporting, PP4_Moderate, PP1_Moderate, PM2_Supporting.