Lysosomes pp 265-291 | Cite as

Prenatal Diagnosis of Lysosomal Storage Disorders Using Chorionic Villi

  • Jyotsna Verma
  • Sunita Bijarnia-MahayEmail author
  • Ishwar C. Verma
Part of the Methods in Molecular Biology book series (MIMB, volume 1594)


Prenatal enzymatic diagnosis for an array of lysosomal storage disorders (LSDs) can be performed accurately, provided that a confirmed diagnosis by biochemical/molecular study in the index case is available and a strict defined protocol, specific to each individual disorder is followed. The present chapter describes the protocols for reliable and accurate prenatal enzymatic diagnoses by fluorometric and spectrophotometric methods of lysosomal storage disorders: Gaucher, Fabry, Pompe, Niemann Pick A/B, Tay Sach, Sandhoff, GM1, Mucoplysaccharidoses, Wolman, Krabbe, Metachromatic leukodystrophy, and Batten diseases using uncultured chorionic villi samples. The biological reference intervals for enzyme levels in normal and affected fetuses are given for interpretation of prenatal results. It is imperative to establish normal reference interval in each laboratory to take into account the local environment, technical variations, and different ethnicities. Besides, enzyme activity in the fetus should be represented as percentage of the mean activity of enzyme of normal fetuses. The pitfalls and challenges in prenatal diagnosis as well as technical problems in performing enzyme assays are also discussed to help the reader in standardization and performing the assays for correct diagnosis.

Key words

Prenatal diagnosis Lysosomal storage disorders Enzymatic diagnosis Chorionic villi Reference intervals 



We would like to thank our colleagues Ratna D. Puri for clinical suggestions, Renu Saxena and Sudha Kohli for their cooperation in molecular analysis, Divya C. Thomas for scientific discussion, Sandeepika, Geetu and Azad for technical assistance and the Chairman of the Hospital for his support in setting up the genetic facility.


  1. 1.
    Verma J, Thomas DC, Sharma S (2015) Inherited metabolic disorders: prenatal diagnosis of lysosomal storage disorders. Prenat Diagn 35:1–11CrossRefGoogle Scholar
  2. 2.
    Sheth J, Sheth F (2013) Genetics in clinical practice—symptoms, diagnosis and therapy. Jaypee Brothers, Ahmedabad, Gujarat. ISBN: 978-93-5152-153-2Google Scholar
  3. 3.
    Fletcher JM (2006) Screening for lysosomal storage disorders: a clinical perspective. J Inherit Metab Dis 29(2–3):405–408CrossRefPubMedGoogle Scholar
  4. 4.
    Beck M (2010) Therapy for lysosomal storage disorders. IUBMB Life 62:33–40PubMedGoogle Scholar
  5. 5.
    Kaminsky P, Lidove O (2014) Current therapeutic strategies in lysosomal disorders. Presse Med 43(11):1174–1184CrossRefPubMedGoogle Scholar
  6. 6.
    Winchester B, Young E (1991) Prenatal diagnosis of enzyme defect—an update. Arch Dis Child 66:451–454CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Verma J, Thomas DC, Sharma S et al (2015) Inherited metabolic disorders: quality management in laboratory diagnosis. Clin Chim Acta 447:1–7CrossRefPubMedGoogle Scholar
  8. 8.
    Verma J, Thomas DC, Verma IC (2016) New generation techniques in rapid diagnosis of lysosomal storage disorders. Eur J Mol Biol Biochem 3(1):42–47Google Scholar
  9. 9.
    Peter SP, Lee RE, Glew RH (1975) A micro assay for Gaucher disease. Clin Chim Acta 60:391–396CrossRefGoogle Scholar
  10. 10.
    Daniels LB, Glew RH (1982) Beta glucosidase assays in the diagnosis of Gaucher disease. Clin Chem 28:569–577PubMedGoogle Scholar
  11. 11.
    Arora D, Kochar SPS (2010) Prenatal diagnosis of Gaucher disease. Med J Armed Forces India 66:170–171CrossRefPubMedGoogle Scholar
  12. 12.
    van Diggelen OP, Voznyi YV, Keulemans JLM et al (2005) A new fluorimetric enzyme assay for the diagnosis of Niemann–Pick A/B, with specificity of natural sphingomyelinase substrate. J Inherit Metab Dis 28(5):733–741CrossRefPubMedGoogle Scholar
  13. 13.
    Suzuki K, Glew R, Peters SP (1977) Practical enzymology of the sphingolipidosis. AR Liss, New York, pp 101–136Google Scholar
  14. 14.
    Grabowski GA, Kruse JR, Goldberg JD et al (1984) First-trimester prenatal diagnosis of Tay-Sachs disease. Am J Hum Genet 36(6):1369–1378PubMedPubMedCentralGoogle Scholar
  15. 15.
    Percy AK, Brady RO (1968) Metachromatic leukodystrophy: diagnosis with samples of venous blood. Science 161(3841):594–595CrossRefPubMedGoogle Scholar
  16. 16.
    Galjaard H, van Hoogstraten JJ, de Jong JE et al (1974) Methodology of the quantitative cytochemical analysis of single or small numbers of cultured cells. Histochem J 6(4):409–429CrossRefPubMedGoogle Scholar
  17. 17.
    Giles L, Coope A, Fowler B et al (1987) Aryl sulfatase isoenzymes of chorionic villi: implications for prenatal diagnosis. Prenat Diagn 7(4):245–252CrossRefPubMedGoogle Scholar
  18. 18.
    Wiederschain G, Raghvan S, Kolodny E (1992) Characterization of 6-hexadecanoylamino-4-methylumbelliferyl-beta-D-galactopyranoside as fluorogenic substrate of galactocerebrosidase for the diagnosis of Krabbe disease. Clin Chim Acta 205(1–2):87–96CrossRefPubMedGoogle Scholar
  19. 19.
    PrajnyaR RC, Phadke SR et al (2011) Prenatal diagnosis of Pompe disease—enzyme assay or molecular testing? Indian Pediatr 48(11):901–902CrossRefGoogle Scholar
  20. 20.
    Chamoles NA, Blanco M, Gaggioli D (2001) Fabry disease: enzymatic diagnosis in dried blood spots on filter paper, letter to the editor. Clin Chim Acta 308:195–196CrossRefPubMedGoogle Scholar
  21. 21.
    Van Diggelen OP, Keulemans JLM, Winchester B et al (1999) A rapid fluorogenic palmitoyl protein thioesterase assay: pre and post natal diagnosis of INCL. Mol Genet Metab 66:240–244CrossRefPubMedGoogle Scholar
  22. 22.
    Hopwood JJ, Muller V, Smithson A et al (1979) A flourometric assay using 4 methylumbelliferyl alpha-l-iduronide for the estimation of alpha-l-iduronidase activity and the detection of Hurler and Schie syndromes. Clin Chem Acta 92(2):257–265CrossRefGoogle Scholar
  23. 23.
    Voznyi YV, Keulemans JLM, van Digglen OP (2001) A flourogenic assay for the diagnosis of hunter disease (MPS II). J Inherit Metab Dis 24:675–680CrossRefPubMedGoogle Scholar
  24. 24.
    Karpova EA, Voznyi YV, Keulemans JLM et al (1996) A fluorometric assay for the diagnosis of Sanfilippo disease type A (MPS IIIA). J Inherit Metab Dis 19:278–285CrossRefPubMedGoogle Scholar
  25. 25.
    Marsh J, Fensom AH (1985) 4-Methylumbelliferyl-α-N-acetylglucosaminidase activity for diagnosis of Sanfilippo B disease. Clin Genet 27:258–262CrossRefPubMedGoogle Scholar
  26. 26.
    Zao H, van Diggelen OP, Thoomes R et al (1990) Prenatal diagnosis of Morquio disease type A using a simple fluorometric enzyme assay. Prenat Diagn 10:85–91CrossRefGoogle Scholar
  27. 27.
    van Diggelen OP, Zhao H, Kleijer WJ et al (1990) A fluorometric enzyme assay for the diagnosis of Morquio disease type A. Clin Chem Acta 187:131–140CrossRefGoogle Scholar
  28. 28.
    Kally S (1977) Biochemical methods in medical genetics. American lecture series 1977; Publication no. 1008. Harvard, 18th edn. Springfield III, Thomas, ChicagoGoogle Scholar
  29. 29.
    Sanguinetti N, Marsh J, Jackson M et al (1986) The aryl sulfatases of chorionic villi: potential problems in the first trimester diagnosis of Metachromatic leukodystrophy and Maroteaux-lamy disease. Clin Genet 30(4):302–308CrossRefPubMedGoogle Scholar
  30. 30.
    Patrick D, Willcox P, Stephens R et al (1976) Prenatal diagnosis of Wolman’s disease. J Med Genet 13:49–51. doi: 10.1136/jmg.13.1.49 CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Huggins C, Lapides J (1947) Chromogenic substrates IV. Acyl esters of p-nitrophenol as substrates for the colorimetric determination of esterase. J Biol Chem 170:467–482Google Scholar
  32. 32.
    Lowry OH, Rosebrough NJ, Farr AL et al (1951) Determination of protein concentration. J Biol Chem 193:265–275PubMedGoogle Scholar
  33. 33.
    Fowler B, Giles L, Cooper A et al (1989) Chorionic villi sampling: diagnostic uses and limitations of enzyme assays. J Inherit Metab Dis 12 suppl(1):105–117Google Scholar
  34. 34.
    Verma J, Thomas DC, Kasper DC et al (2016) Inherited metabolic disorders: efficacy of enzyme assays on dried blood spots for the diagnosis of lysosomal storage disorders. JIMD Rep. doi: 10.1007/8904_2016_548 PubMedPubMedCentralGoogle Scholar
  35. 35.
    Miller SA, Dykes DD, Polesky HF (1988) A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 16(3):1215CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Sheth J, Mistri M, Sheth F et al (2014) Prenatal diagnosis of lysosomal storage disorders by enzymes study using chorionic villus and amniotic fluid. J Fetal Med 1:17–24CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Jyotsna Verma
    • 1
  • Sunita Bijarnia-Mahay
    • 1
    Email author
  • Ishwar C. Verma
    • 1
  1. 1.Institute of Medical Genetics and Genomics, Sir Ganga Ram HospitalNew DelhiIndia

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