Advertisement

Journal of Inherited Metabolic Disease

, Volume 33, Issue 1, pp 43–50 | Cite as

Diagnostic efficacy of the fluorometric determination of enzyme activity for Pompe disease from dried blood specimens compared with lymphocytes—possibility for newborn screening

  • Zoltan Lukacs
  • Paulina Nieves Cobos
  • Eugen Mengel
  • Ralf Hartung
  • Michael Beck
  • Marcus Deschauer
  • Angelika Keil
  • René Santer
Original Article

Abstract

Background

Pompe disease is a rare, autosomal-recessive disorder which results from a defect in the lysosomal enzyme acid α-glucosidase (GAA). The onset of this disease is highly variable, with infantile types being the most severe. Traditionally, lymphocytes, fibroblasts or muscle biopsies were necessary for enzyme activity measurement, because these materials do not express maltase-glucoamylase (MGA) that interferes with the assay. Recently, acarbose was found to inhibit MGA activity selectively, so that dried blood became accessible for GAA assessment.

Aim

To evaluate the diagnostic efficacy of GAA measurement in dried blood specimens (DBSs) in comparison with lymphocytes. If DBSs provided reliable results, the diagnosis of Pompe disease could be facilitated, and high-throughput screening would become possible.

Methods and results

GAA activity was measured in DBSs of known patients at pH 3.8 (with and without acarbose) and at pH 7.0. Additionally, lymphocytes were obtained from the same patients, and the enzyme activity was determined at pH 4 to pH 7. In total, seven infantile patients and 29 patients with late-onset variants were investigated. All patients were reliably identified by both methods. Furthermore, a simplified protocol was established for neonatal screening.

Conclusion

The fluorometric technique for the assessment of GAA activity in DBS provides a reliable diagnosis for all variants of Pompe disease. The assay protocol could be simplified for neonatal screening, without increasing the false positive rate significantly or burdening the laboratory with time-consuming procedures.

Keywords

Enzyme Replacement Therapy Acarbose Pompe Disease Neonatal Screening Pompe Patient 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

The study was supported by the Genzyme Corp., USA. We also wish to express our gratitude to Dr. Joan Keutzer for her continuous help and support and to Dr. Gläser (Neu-Ulm) for assisting in the molecular genetic characterization of some patients.

References

  1. Baethmann M, Straub V, Reuser AJJ (eds) (2008) Pompe disease. UNI-MED, BremenGoogle Scholar
  2. Bembi B, Cerini E, Danesino C et al (2008) Management and treatment of glycogenosis type II. Neurology 71(Suppl 2):S12–S36CrossRefPubMedGoogle Scholar
  3. Chamoles N, Niizawa G, Blanco M, Gaggioli D, Casentini C (2004) Glycogen storage disease type II: enzymatic screening in dried blood spots on filter paper. Clin Chim Acta 347:97–102CrossRefPubMedGoogle Scholar
  4. Chien YH, Chiang SC, Zhang XK, et al (2008) Early detection of Pompe disease by newborn screening is feasible: results from the Taiwan screening program. Pediatrics 122:39–45CrossRefGoogle Scholar
  5. Dajnoki A, Mühl A, Fekete G et al (2008) Newborn screening for Pompe disease by measuring acid α-glucosidase activity using tandem mass spectrometry. Clin Chem 54:1624–1629CrossRefPubMedGoogle Scholar
  6. Dreyfus J, Poenaru L (1980) White blood cells and the diagnosis of alpha-glucosidase deficiency. Pediatr Res 14:342–344CrossRefPubMedGoogle Scholar
  7. Jack RM, Gordon C, Scott CR, Kishnani PS, Bali D (2006) The use of acarbose inhibition in the measurement of acid alpha-glucosidase activity in blood lymphocytes for the diagnosis of Pompe disease. Genet Med 8:307–312CrossRefPubMedGoogle Scholar
  8. Joshi PR, Gläser D, Schmidt S et al (2008) Molecular diagnosis of German patients with late-onset glycogen storage disease type II. J Inherit Metab Dis. doi: 10.1007/s10545-008-0820-2
  9. Kishnani PS, Steiner RD, Bali D et al (2006a) Pompe disease diagnosis and management guideline. Genet Med 8:267–288CrossRefGoogle Scholar
  10. Kishnani PS, Hwu WL, Mandel H et al (2006b) A retrospective, multinational, multicenter study on the natural history of infantile-onset Pompe disease. J Pediatr 148:671–676CrossRefGoogle Scholar
  11. Kumamoto S, Katafuchi T, Nakamura K et al (2009) High frequency of acid alpha-glucosidase pseudodeficiency complicates newborn screening for glycogen storage disease type II in the Japanese population. Mol Genet Metab 97:190–195CrossRefPubMedGoogle Scholar
  12. Lowry OH, Roseborough NJ, Farr AL et al (1951) Protein measurements with Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  13. Müller-Felber W, Horvath R, Gempel K et al (2007) Late-onset Pompe disease: clinical and neurophysiological spectrum of 38 patients including long-term follow-up in 18 patients. Neuromuscul Disord 17:698–706CrossRefPubMedGoogle Scholar
  14. Okumiya T, Keulemans JLM, Kroos MA et al (2006) A new diagnostic assay for glycogen storage disease type II in mixed leukocytes. Mol Genet Metab 88:22–28CrossRefPubMedGoogle Scholar
  15. Ploeg AT, Reuser AJJ (2008) Lysosomal storage disease 2 Pompe’s disease. Lancet 372:1342–1353CrossRefPubMedGoogle Scholar
  16. Winchester B, Bali D, Bodamer OA et al (2008) Methods for a prompt and reliable laboratory diagnosis of Pompe disease: report from an international consensus meeting. Mol Genet Metab 93:275–281CrossRefPubMedGoogle Scholar
  17. Zhang H, Kallwass H, Young SP et al (2006) Comparison of maltose and acarbose as inhibitors of maltase-glucoamylase activity in assaying acid α-glucosidase activity in dried blood spots for the diagnosis of infantile Pompe disease. Genet Med 8:302–306CrossRefPubMedGoogle Scholar

Copyright information

© SSIEM and Springer 2009

Authors and Affiliations

  • Zoltan Lukacs
    • 1
    • 2
    • 5
  • Paulina Nieves Cobos
    • 1
    • 2
  • Eugen Mengel
    • 3
  • Ralf Hartung
    • 3
  • Michael Beck
    • 3
  • Marcus Deschauer
    • 4
  • Angelika Keil
    • 1
    • 2
  • René Santer
    • 2
  1. 1.Institute of Clinical ChemistryUniversity Medical Center of Hamburg-EppendorfHamburgGermany
  2. 2.Department of PediatricsUniversity Medical Center of Hamburg-EppendorfHamburgGermany
  3. 3.Department of PediatricsUniversity of MainzMainzGermany
  4. 4.Department of NeurologyUniversity of Halle-WittenbergHalleGermany
  5. 5.Metabolic Laboratory/Institute of Clinical ChemistryUniversity Medical Center of Hamburg-EppendorfHamburgGermany

Personalised recommendations