To study 160 patients with undiagnosed LGMD, we first sequenced the hotspot regions at exons 5 and 20 of the ANO5 gene, but the screening was diagnostic in two families only (I and XVI, homozygotes for c.191dupA and c.2272 C > T, respectively). The gene discussed is ANO5; the disease is limb-girdle muscular dystrophy.