Journal of Inherited Metabolic Disease

, Volume 32, Issue 3, pp 416–423 | Cite as

Enzyme analysis for Pompe disease in leukocytes; superior results with natural substrate compared with artificial substrates

  • O. P. van Diggelen
  • L. F. Oemardien
  • N. A. M. E. van der Beek
  • M. A. Kroos
  • H. K. Wind
  • Y. V. Voznyi
  • D. Burke
  • M. Jackson
  • B. G. Winchester
  • A. J. J. Reuser
Original Article


Enzyme analysis for Pompe disease in leukocytes has been greatly improved by the introduction of acarbose, a powerful inhibitor of interfering α-glucosidases, which are present in granulocytes but not in lymphocytes. Here we show that the application of acarbose in the enzymatic assay employing the artificial substrate 4-methylumbelliferyl-α-d-glucoside (MU-αGlc) is insufficient to clearly distinguish patients from healthy individuals in all cases. Also, the ratios of the activities without/with acarbose only marginally discriminated Pompe patients and healthy individuals. By contrast, when the natural substrate glycogen is used, the activity in leukocytes from patients (n = 82) with Pompe disease is at most 17% of the lowest control value. The use of artificial substrate in an assay with isolated lymphocytes instead of total leukocytes is a poor alternative as blood samples older than one day invariably yield lymphocyte preparations that are contaminated with granulocytes. To diagnose Pompe disease in leukocytes we recommend the use of glycogen as substrate in the presence of acarbose. This assay unequivocally excludes Pompe disease. To also exclude pseudo-deficiency of acid α-glucosidase caused by the sequence change c.271G>A (p.D91N or GAA2; homozygosity in approximately 1:1000 caucasians), a second assay employing MU-αGlc substrate plus acarbose or DNA analysis is required.


Acarbose Glycogen Storage Disease Pompe Disease Glcn 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.



dried blood spot on filter paper


glycogen storage disease type II, Pompe disease, acid maltase deficiency











We thank Ton de Wit for communicating the cytometry experiments.

This study was supported by a grant from Genzyme Corporation, Cambridge MA, USA and was part of the Dutch TI Pharma initiative to commence a project on Sustainable Orphan Drug Development through Registries and Monitoring (T6–208).


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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • O. P. van Diggelen
    • 1
    • 8
  • L. F. Oemardien
    • 1
  • N. A. M. E. van der Beek
    • 2
  • M. A. Kroos
    • 1
  • H. K. Wind
    • 3
  • Y. V. Voznyi
    • 4
  • D. Burke
    • 5
  • M. Jackson
    • 6
  • B. G. Winchester
    • 7
  • A. J. J. Reuser
    • 1
  1. 1.Department of Clinical GeneticsErasmus University Medical CentreRotterdamThe Netherlands
  2. 2.Department of NeurologyErasmus University Medical CentreRotterdamThe Netherlands
  3. 3.Department of ImmunologyErasmus University Medical CentreRotterdamThe Netherlands
  4. 4.Institute of Organic ChemistryMoscowRussia
  5. 5.The Enzyme Laboratory, Department of Chemical PathologyGreat Ormond Street HospitalLondonUK
  6. 6.Supra-regional Assay Service for Genetic Enzyme DefectsGuys HospitalLondonUK
  7. 7.Biochemistry Research Group, UCL Institute of Child HealthUniversity College LondonLondonUK
  8. 8.Department of Clinical GeneticsErasmus MCRotterdamThe Netherlands

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