Use of a multiplex ligation-dependent probe amplification method for the detection of deletions/duplications in the GBA1 gene in Gaucher disease patients.
Gaucher disease (GD) is caused by the deficient activity of β-glucocerebrosidase due to pathogenic mutations in the GBA1. This gene has a pseudogene (GBAP) with 96% of sequence homology. Recombination (Rec) events in the GBA1 seem to be facilitated by an increased degree of homology and proximity to the GBAP. The objectives of this study were to validate the P338-X1 GBA kit (MRC-Holland) for Multiplex Ligation-dependent Probe Amplification (MLPA) and to detect larger deletions/duplications present in GBA1 in GD patients from Brazil. Thirty-three unrelated Brazilian GD patients, previously genotyped by the Sanger method (both pathogenic alleles identified=29 patients, only one allele identified=3 patients, no pathogenic alleles identified=1 patient), were evaluated by the MLPA assay. MLPA was compatible with the previous results obtained by Sanger sequencing and identified an additional allele (a heterozygous deletion in intron 7 in one patient with only one mutation identified by Sanger). Our data suggest that, although larger deletions/duplications do not appear to be frequent in GD, the P338-X1 GBA kit for MLPA appears to be a good method for GBA1 analysis. Additional investigations should be performed in order to characterize the remaining four uncharacterized alleles of our sample.
Postgraduate Program in Medical Sciences, School of Medical Sciences, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Basic Research and Advanced Investigations in Neurosciences (BRAIN) Laboratory, Experimental Research Center, Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, Brazil.
Suelen P
SP
Basgalupp
Postgraduate Program in Sciences of Gastroenterology and Hepatology, School of Medical Sciences, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Basic Research and Advanced Investigations in Neurosciences (BRAIN) Laboratory, Experimental Research Center, Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, Brazil; Molecular and Protein Analysis Unit, Experimental Research Center, Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, Brazil.
Marina
M
Siebert
Medical Genetics Service, Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, Brazil.
Filippo Pinto E
FPE
Vairo
Rede Mater Dei de Saúde, Belo Horizonte, Brazil.
Anisse Marques
AM
Chami
Hospital Infantil Joana de Gusmão, Florianópolis, Brazil.
Louise Lapagesse de Camargo
LLC
Pinto
Hospital da Criança de Brasilia José de Alencar; Brasília, Brazil.
Gerson da S
GDS
Carvalho
Postgraduate Program in Medical Sciences, School of Medical Sciences, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Basic Research and Advanced Investigations in Neurosciences (BRAIN) Laboratory, Experimental Research Center, Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, Brazil; Medical Genetics Service, Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, Brazil; Department of Genetics, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil. Electronic address: idadschwartz@gmail.com.
Ida Vanessa D
IVD
Schwartz
D000483:Alleles; D001938:Brazil / Q000453:epidemiology; D005091:Exons; D005776:Gaucher Disease / Q000175:diagnosis / Q000453:epidemiology / Q000235:genetics*; D017353:Gene Deletion; D020440:Gene Duplication; D005838:Genotype; D005962:Glucosylceramidase / Q000235:genetics*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D017354:Point Mutation
D005962:Glucosylceramidase
GBA1; Gaucher disease; MLPA; Molecular diagnosis