Enzymological Laboratory

The lysosomal enzyme laboratory provides a national and international diagnostic service for the lysosomal storage disorders (LSD). Enzyme diagnosis focused on a group of sfingolipidoses started in the Czech Republic in the mid eighties of the last century. The spectrum of the enzyme diagnostics has grown to virtually all known lysosomal enzymopathies over the last 20 years and currently offers postnatal and prenatal diagnoses for over 40 different disorders from the group of sfingolipideses, glykoproteinoses, mukopolysaccharidoses and some other rare diseases (mucolipidoses, Wolman disease and CESD, deficiences of saposin activators). In the past three decades, a total of 556 (93 Fabry heterozygots not included) patients have been diagnosed with one of 34 specific types of LSDs in the Czech Republic.

The lysosomal enzyme panel tests for a range of lysosomal storage disorders (see bellow) can be performed either on leukocytes of peripheral blood or on cultured cells (skin fibroblasts) mostly using synthetic analogues of natural enzyme substrates. For screening studies of some LSD, enzyme analysis in dry blood spots has been introduced (Pompe disease, Fabry disease). Some lysosomal hydrolases can be screened also in the blood serum or plasma (ML II/III, Tay-Sachs and Sandhoff diseases). For prenatal diagnosis, chorionic villi or cultivated amniocytes are used. Chromatographic or MS / MS analysis of sphingolipids in urine or in tissue samples is provided as a useful supplementary or screening test.

In close association with other groups, enzyme laboratory is also engaged in different research projects focused on pathobiochemistry and cell biology of LSD and the development of diagnostic methods.

Lysosomal laboratory is accredited according to CSN EN ISO 15189:2013 and is a member of the European Study Group of Lysosomal Disorders and since 2006 is involved in the international system of external quality control QA-ESGLD/ERDNIM.

List of Methods

Disorder Material Enzyme/Metabolite
Disorders of biogenic amine metabolism
Tetrahydrobiopterin (BH4) deficiency dried blood spot dihydropteridine reductase
Disorders of carbohydrate metabolism - Disorder of galactose metabolism
Galactosemia dried blood spot galactose-1-phosphate uridyltransferase
Disorders of purine metabolism
Lesch-Nyhan syndrome erythrocytes hypoxanthine:guanine phosphoribosyltransferase
Adenine phosphoribosyltransferase deficiency erythrocytes adenine phosphoribosyltransferase
Adenosine deaminase deficiency erythrocytes adenosine deaminase
Phosphoribosyl pyrophosphate synthetase superactivity erythrocytes phosphoribosyl pyrophosphate synthetase
Purinnenucleosidephosphorylase deficiency erythrocytes purine nucleoside phosphorylase
Disorders of carbohydrate metabolism - Glycogen storage disorders
Glycogen storage disease type II (Pompe) leukocytes, dried blood spot, fibroblasts acid α-1,4-glucosidase
Glycogen storage disease type IV erythrocytes amylo-1,4,-1,6-transglukosidosis
Glycogen storage disease type VI leucocytes liver phosphorylase
Glycogenos storage disease type XI erythrocytes phosphorylase-b-kinase
Lysosomal disorders - Screening
Lysosomal storage diseases plasma, serum chitotriosidase
Metachromatic leukodystrophy (MLD) urine sulfatides
Lysosomal disorders - Mucopolysaccharidoses
Mucopolysaccharidoses type I (Hurler, Scheile) leukocytes, fibroblasts α-L-iduronidase
Mucopolysaccharidoses type II (Hunter) leukocytes, plasma, serum, fibroblasts iduronate sulphatase
Mucopolysaccharidoses type III A (m. Sanfillipo A) leukocytes, fibroblasts heparinsulphamidase
Mucopolysaccharidoses type III B (m. Sanfillipo B) leukocytes, plasma, serum, fibroblasts α-N-acetyl-D-glucosaminidase
Mucopolysaccharidoses type III C (m. Sanfillipo C) leukocytes, fibroblasts acetyl-CoA:α-glucosaminide N-acetyltransferase
Mucopolysaccharidoses type III D (m. Sanfillipo D) leukocytes, fibroblasts N-acetylglucosamine-6-sulphate-sulphatase
Mucopolysaccharidoses type IV A (m. Morquio A) leukocytes, fibroblasts N-acetylgalactosamine-6-sulphate-sulphatase
Mucopolysaccharidoses type IV B (m. Morquio B) leukocytes, fibroblasts β-galactosidase
Mucopolysaccharidoses typeVI (Maroteaux-Lamy) leukocytes, fibroblasts arylsulfatase B (N-acetylgalactosamine-4-sulphatase)
Mucopolysaccharidoses typeVII (Sly) leukocytes, fibroblasts β-glucuronidase
Lysosomal disorders - Sphingolipidoses and lipidoses
GM1 Gangliosidosis leukocytes, fibroblasts β-galactosidase
Metachromatic leukodystrophy (MLD) leukocytes, fibroblasts arylsulfatase A
Gaucher disease leukocytes, fibroblasts glucocerebrosidase (acid β-glucosidase)
Fabry disease leukocytes, plasma, serum, fibroblasts, dried blood spot (males) α-galactosidase
Krabbe disease leukocytes, fibroblasts galactocerebrosidase
Niemann-Pick disease type I (= type A, B) leukocytes, fibroblasts acid sphingomyelinase
GM2 Gangliosidosis (Sandhoff disease) leukocytes, plasma, serum, fibroblasts β-hexosaminidase
GM2 Gangliosidosis (Tay-Sachs disease) leukocytes, plasma, serum, fibroblasts β-hexosaminidase A
Wolman disease leukocytes, fibroblasts acid lipase
Prosaposin deficiency (pSap-d) and saposin B deficiency (SapB-d) urine glycolipids
Lysosomal disorders -Oligosaccharidoses
α-Mannosidosis leukocytes, fibroblasts α-mannosidase
β-Mannosidosis leukocytes, fibroblasts β-mannosidase
Fucosidosis leukocytes, fibroblasts α-L-fucosidase
Schindler disease leukocytes, fibroblasts α-N-acetyl-D-galactosaminidase
Sialidosis fibroblasts α-neuraminidase
Lysosomal disorders - Mucolipidoses
Mukolipidoses II, III, I-cell disease plasma, serum, fibroblasts N-acetyl-glucosamine-1-phosphotransferase
Lysosomal disorders - Ceroid lipfuscinoses, neuronal (CLN)
Neuronal ceroid lipofuscinose type 1 (NCL I) leukocytes, fibroblasts palmitoyl-proteinthioesterase
Neuronal ceroid lipofuscinose type 2 (NCL II) leukocytes, fibroblasts tripeptidylpeptidase I
Other disorders
X-linked ichthyosis leukocytes, fibroblasts steroidsulphatase, arylsulfatase C
Biotinidase deficiency dried blood spot, serum biotinidase