Ultramicro-fluorometric assay for the diagnosis of Gaucher disease in dried blood spots on filter paper.
J Neonatal Perinatal Med
; 6(1): 61-7, 2013.
Article
em En
| MEDLINE
| ID: mdl-24246460
BACKGROUND: Gaucher disease (GD) is a lysosomal storage disorder characterized by a deficiency of the lysosomal acid ß-D-glucosidase (GBA). The aim of this study was to develop an ultramicro-fluorometric assay based on the method of Chamoles et al. for determining GBA activity in dried blood spots on filter paper (DBS). METHODS: The assay used 3-mm diameter blood spot and 8 mmol/l of 4-methylumbelliferyl-ß-D-glucoside as enzymatic substrate. The reaction occurred in plates incubated at 37°C for 20 hours and the enzyme activity was expressed in µmol hydrolysed substrate/l blood/h. The fluorescence of the enzyme product was automatically measured in a fluorometer-photometer reader (SUMA Technology). RESULTS: The intra and inter-assay coefficients of variation were lower than 9 and 12%, respectively, and the recovery range was 97-109%.Three patients with GD were correctly diagnosed using the ultramicroassay. Healthy newborn DBS samples (n = 3003) from the National Neonatal Screening Program were analyzed, and the mean GBA activity was 5.7 µmol/l blood/h. Our assay showed high Pearson (n = 26; r = 0.99) and concordance correlations (ρc = 0.99) with the traditional method described by Chamoles et al. CONCLUSIONS: The analytical performance characteristics of our ultramicro-fluorometric assay suggest that it can be used in the diagnosis of GD in newborns and adults.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Triagem Neonatal
/
Teste em Amostras de Sangue Seco
/
Fluorometria
/
Doença de Gaucher
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
Limite:
Female
/
Humans
/
Male
/
Newborn
País/Região como assunto:
Caribe
/
Cuba
Idioma:
En
Revista:
J Neonatal Perinatal Med
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
Cuba
País de publicação:
Holanda