The NM_213599.3: c.1295C>G variant in ANO5 is a missense variant predicted to cause the substitution of alanine by glycine at amino acid position 432, p.(Ala432Gly). The computational predictor REVEL gives a score of 0.176, however, SpliceAI gives a delta score of 0.62 for loss of the canonical donor and 1.00 for gain of a cryptic donor 38 nucleotides into exon 13. RNA analysis has demonstrated a complete splice effect of this variant, resulting in the deletion of the last 38 nucleotides of exon 13, which is expected to lead to a frameshift, premature truncation, p.(Ala432GlyfsTer49), and subsequent nonsense mediated decay (PMID: 20096397, PVS1_RNA). This variant has been reported in at least five unrelated individuals with features consistent with LGMD (PMID: 26809617, 31353849, 20096397), including in a homozygous state in the probands of two consanguineous families sharing a common haplotype (0.25 pts x2; PMID: 20096397) and in unknown phase with a pathogenic variant in three affected individuals (c.989dupT p.(Leu330Phefs), 0.5 pts, PMID: 26809617; c.191dupA p.(Asn64LysfsTer15), 0.5 pts, PMID: 31353849; and c.2272C>T p.(Arg758Cys), 0.5 pts, PMID: 31353849) (PM3_Strong). At least one patient with this variant and a second presumed diagnostic ANO5 allele displayed a progressive limb girdle pattern of muscle weakness (PMID: 26809617, 31353849, 20096397; PP4). This variant has also been reported to co-segregate with autosomal recessive LGMD in four affected family members from the two families sharing the same haplotype (PMID: 20096397; PP1_Strong). The upper bound of the 95% confidence interval of the Grpmax variant allele frequency in gnomAD v4.1.0 is 0.000021617 (17/1179582 European (non-Finnish) chromosomes), which is less than the threshold of 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 2.0.0; 01/08/2026): PVS1_RNA, PM3_Strong, PP4, PP1_Strong, PM2_Supporting.