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 |
