title: A framework for the development of STR genotyping in domestic animal species: characterization and population study of 12 canine X-chromosome loci.
abstract: This study reports the methodology used to search, select and characterize STR loci on the canine X chromosome using publicly available genome resources and following the current guidelines for human and non-human forensic testing. After several rounds of selection, 12 X-STR markers were optimized for simultaneous co-amplification in a single PCR, and genetic profiles were determined in a sample of 103 unrelated dogs. Mendelian inheritance was verified and mutation rates were assessed using family groups. Alleles that varied in size were sequenced to create a standardized nomenclature proposal based on the number of repeats. All loci conformed to Hardy-Weinberg expectations. The resulting panel showed high forensic efficiency, presenting high values of power of discrimination (in males and females) and mean exclusion chance, both in trios involving female offspring and in duos composed of dam and male offspring. Its use may complement the information obtained by autosomal STR analysis and contribute to the resolution of complex cases of kinship in dogs. The presented methodology for the de novo construction of an STR multiplex may also provide a helpful framework for analogous work in other animal species. As an increasing number of reference genomes become available, convenient tools for individual identification and parentage testing based on STR loci selected from autosomes or sex chromosomes' sequences may be created following this strategy.
authors.affiliation: Instituto de Patologia e Imunologia da Universidade do Porto, Porto, Portugal. basch@ipatimup.pt
authors.forename: Barbara
authors.initials: B
authors.lastname: van Asch
authors.forename: Raquel
authors.initials: R
authors.lastname: Pinheiro
authors.forename: Rui
authors.initials: R
authors.lastname: Pereira
authors.forename: Cíntia
authors.initials: C
authors.lastname: Alves
authors.forename: Vânia
authors.initials: V
authors.lastname: Pereira
authors.forename: Filipe
authors.initials: F
authors.lastname: Pereira
authors.forename: Leonor
authors.initials: L
authors.lastname: Gusmão
authors.forename: António
authors.initials: A
authors.lastname: Amorim
mesh_terms: D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D004285:Dogs / Q000235:genetics*; D005260:Female; D005787:Gene Frequency; D056426:Genetic Loci; D014644:Genetic Variation; D042822:Genomic Instability; D005838:Genotype; D015810:Linkage Disequilibrium; D008297:Male; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D014960:X Chromosome