Chitotriosidase determination in plasma and in dried blood spots: A comparison using two different substrates in a microplate assay

Chitotriosidase (CT) is a macrophage glycosylhydrolase referred as a biochemical marker on diagnosis and prognosis for Gaucher Disease (GD). The aim of the study was to validate a microplate assay for plasma and dried blood spots on filter paper (DBS) for CT activity determination and to compare CT...

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Bibliographic Details
Published in:Clinica chimica acta 2009-08, Vol.406 (1), p.86-88
Main Authors: Rodrigues, Mayra D.B., de Oliveira, Allan C., Müller, Karen B., Martins, Ana Maria, D'Almeida, Vânia
Format: Article
Language:English
Subjects:
Adolescent
Adult
Blood Chemical Analysis - methods
Case-Control Studies
Child
Chitotriosidase
Dried blood spot
Female
Gaucher disease
Gaucher Disease - blood
Gaucher Disease - diagnosis
Gaucher Disease - therapy
Hexosaminidases - blood
Hexosaminidases - metabolism
Humans
Male
Middle Aged
Trisaccharides - chemistry
Trisaccharides - metabolism
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Summary:Chitotriosidase (CT) is a macrophage glycosylhydrolase referred as a biochemical marker on diagnosis and prognosis for Gaucher Disease (GD). The aim of the study was to validate a microplate assay for plasma and dried blood spots on filter paper (DBS) for CT activity determination and to compare CT activity using 4-methylumbelliferyl-b-D-N,N′,N″-triacetyl-chitotrioside (4MU-C3) and 4-methylumbelliferyl-deoxychitobiose (4MU-dC2) as substrates. Heparinized blood was collected from 12 healthy volunteers (HV) and 14 treated GD patients. Total blood was used to prepare DBS and plasma was obtained by centrifugation. CT activity measurements were done on tube and microplate by a fluorimetric assay using plasma and punches of the DBS samples. The use of 4MU-C3 presented a good correlation among plasma and DBS when analyzing the groups altogether (Pearson = 0.76), but the correlation was weaker when we isolated GD patients (Pearson = 0.67). The use of 4MU-dC2 increased the correlation among the samples for this group of individuals (Pearson = 0.89). The possibility of DBS assay facilitates sample transport and storage and CT activity determination on a microplate assay improves laboratory routine. The use of 4MU-dC2 may be more appropriate for GD patients diagnosis on DBS.
ISSN:0009-8981
1873-3492
DOI:10.1016/j.cca.2009.05.022