title: Robust genotyping tool for autosomal recessive type of limb-girdle muscular dystrophies.
abstract: BACKGROUND
  Limb-girdle muscular dystrophies are characterized by predominant involvement of the shoulder and pelvic girdle and trunk muscle groups. Currently, there are 31 genes implicated in the different forms of limb-girdle muscular dystrophies, which exhibit similar phenotypes and clinical overlap; therefore, advanced molecular techniques are required to achieve differential diagnosis.
  
  
  METHODS
  We investigated 26 patients from Latvia and 34 patients from Lithuania with clinical symptoms of limb-girdle muscular dystrophies, along with 565 healthy unrelated controls from general and ethnic populations using our developed test kit based on the Illumina VeraCode GoldenGate genotyping platform, Ion AmpliSeq Inherited Disease Panel and direct sequencing of mutations in calpain 3 (CAPN3), anoctamin 5 (ANO5) and fukutin related protein (FKRP) genes.
  
  
  RESULTS
  Analysis revealed a homozygous CAPN3 c.550delA mutation in eight patients and three heterozygous variants in controls: dysferlin (DYSF) c.5028delG, CAPN3 c.2288A > G, and FKRP c.135C > T. Additionally, three mutations within FKRP gene were found: homozygous c.826C > A, and two compound - c.826C > A/c.404_405insT and c.826C > A/c.204_206delCTC mutations, and one mutation within CLCN1 gene - c.2680C > T p.Arg894Ter. ANO5 c.191dupA was not present.
  
  
  CONCLUSIONS
  Genetic diagnosis was possible in 12 of 60 patients (20%). The allele frequency of CAPN3 gene mutation c.550delA in Latvia is 0.0016 and in Lithuania - 0.0029. The allele frequencies of CAPN3 gene mutation c.2288A > G and DYSF gene mutation c.4872delG are 0.003.
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia. inna@biomed.lu.lv.
authors.forename: Inna
authors.initials: I
authors.lastname: Inashkina
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia.
authors.forename: Eriks
authors.initials: E
authors.lastname: Jankevics
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia.
authors.forename: Janis
authors.initials: J
authors.lastname: Stavusis
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia.
authors.forename: Inta
authors.initials: I
authors.lastname: Vasiljeva
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia.
authors.forename: Kristine
authors.initials: K
authors.lastname: Viksne
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia.
authors.forename: Ieva
authors.initials: I
authors.lastname: Micule
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia.
authors.forename: Jurgis
authors.initials: J
authors.lastname: Strautmanis
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia.
authors.forename: Maruta S
authors.initials: MS
authors.lastname: Naudina
authors.affiliation: Department of Human and Medical Genetics, Faculty of Medicine, Vilnius University, Vilnius, Lithuania.|Centre for Medical Genetics, Vilnius University Hospital Santariškių Klinikos, Santariškių str. 2, LT-08661, Vilnius, Lithuania.
authors.forename: Loreta
authors.initials: L
authors.lastname: Cimbalistiene
authors.affiliation: Department of Human and Medical Genetics, Faculty of Medicine, Vilnius University, Vilnius, Lithuania.
authors.forename: Vaidutis
authors.initials: V
authors.lastname: Kucinskas
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia.
authors.forename: Astrida
authors.initials: A
authors.lastname: Krumina
authors.affiliation: Department of Human and Medical Genetics, Faculty of Medicine, Vilnius University, Vilnius, Lithuania.|Centre for Medical Genetics, Vilnius University Hospital Santariškių Klinikos, Santariškių str. 2, LT-08661, Vilnius, Lithuania.
authors.forename: Algirdas
authors.initials: A
authors.lastname: Utkus
authors.affiliation: Department of Human and Medical Genetics, Faculty of Medicine, Vilnius University, Vilnius, Lithuania.|Centre for Medical Genetics, Vilnius University Hospital Santariškių Klinikos, Santariškių str. 2, LT-08661, Vilnius, Lithuania.
authors.forename: Birute
authors.initials: B
authors.lastname: Burnyte
authors.affiliation: Department of Human and Medical Genetics, Faculty of Medicine, Vilnius University, Vilnius, Lithuania.|Centre for Medical Genetics, Vilnius University Hospital Santariškių Klinikos, Santariškių str. 2, LT-08661, Vilnius, Lithuania.
authors.forename: Ausra
authors.initials: A
authors.lastname: Matuleviciene
authors.affiliation: Biomedical Research and Study Centre, Ratsupites str. 1, k-1, LV-1067, Riga, Latvia.|Laval University, Quebec, Canada.|Centre hospitalier universitaire de Québec, 2705, boulevard Laurier, Québec, Québec, G1V 4G2, Canada.
authors.forename: Baiba
authors.initials: B
authors.lastname: Lace
mesh_terms: D000293:Adolescent; D000328:Adult; D002154:Calpain / Q000235:genetics*; D002648:Child; D002675:Child, Preschool; D015331:Cohort Studies; D005260:Female; D005838:Genotype; D006801:Humans; D007223:Infant; D007844:Latvia / Q000453:epidemiology; D008097:Lithuania / Q000453:epidemiology; D008297:Male; D008875:Middle Aged; D009124:Muscle Proteins / Q000235:genetics*; D049288:Muscular Dystrophies, Limb-Girdle / Q000175:diagnosis* / Q000453:epidemiology / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D055815:Young Adult
chemical_list: D009124:Muscle Proteins; C105884:CAPN3 protein, human; D002154:Calpain
keywords: Calpain 3 c.550delA; Fukutin related protein; Illumina VeraCode GoldenGate; Limb-girdle muscular dystrophies