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. 1991 Feb;28(2):101–109. doi: 10.1136/jmg.28.2.101

Tay-Sachs disease heterozygote detection: use of a centrifugal analyser for automation of hexosaminidase assays with two different artificial substrates.

E C Landels 1, I H Ellis 1, M Bobrow 1, A H Fensom 1
PMCID: PMC1016777  PMID: 1825851

Abstract

An assay for measuring hexosaminidase A in serum and leucocytes is described in which a centrifugal analyser is used for automation of the enzyme assays after manual heat inactivation. The assay was used in a screening programme to identify heterozygotes for Tay-Sachs disease in Ashkenazi Jewish subjects in the UK. The first results from this programme indicate a carrier frequency of 1 in 27. Automation of an assay for direct measurement of hexosaminidase A in serum using 4-methyl-umbelliferyl-beta-N-acetylglucosamine-6-sulphate as substrate is also described. Comparison of data obtained from 66 control and 30 obligate carrier sera tested by this method and by heat inactivation showed improved discrimination using the sulphated substrate. Results obtained using the sulphated substrate for screening serum during pregnancy are also presented.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arpaia E., Dumbrille-Ross A., Maler T., Neote K., Tropak M., Troxel C., Stirling J. L., Pitts J. S., Bapat B., Lamhonwah A. M. Identification of an altered splice site in Ashkenazi Tay-Sachs disease. Nature. 1988 May 5;333(6168):85–86. doi: 10.1038/333085a0. [DOI] [PubMed] [Google Scholar]
  2. Bayleran J., Hechtman P., Saray W. Synthesis of 4-methylumbelliferyl-beta-D-N-acetylglucosamine-6-sulfate and its use in classification of GM2 gangliosidosis genotypes. Clin Chim Acta. 1984 Nov 15;143(2):73–89. doi: 10.1016/0009-8981(84)90215-8. [DOI] [PubMed] [Google Scholar]
  3. Ben-Yoseph Y., Reid J. E., Shapiro B., Nadler H. L. Diagnosis and carrier detection of Tay-Sachs disease: direct determination of hexosaminidase A using 4-methylumbelliferyl derivatives of beta-N-acetylglucosamine-6-sulfate and beta-N-acetylgalactosamine-6-sulfate. Am J Hum Genet. 1985 Jul;37(4):733–740. [PMC free article] [PubMed] [Google Scholar]
  4. Bland J. M., Altman D. G. Statistical methods for assessing agreement between two methods of clinical measurement. Lancet. 1986 Feb 8;1(8476):307–310. [PubMed] [Google Scholar]
  5. Charrow J., Inui K., Wenger D. A. Late onset GM2 gangliosidosis: an alpha-locus genetic compound with near normal hexosaminidase activity. Clin Genet. 1985 Jan;27(1):78–84. doi: 10.1111/j.1399-0004.1985.tb00188.x. [DOI] [PubMed] [Google Scholar]
  6. Dance N., Price R. G., Robinson D., Stirling J. L. Beta-galactosidase, beta-glucosidase and N-acetyl-beta-glucosaminidase in human kidney. Clin Chim Acta. 1969 May;24(2):189–197. doi: 10.1016/0009-8981(69)90311-8. [DOI] [PubMed] [Google Scholar]
  7. Fuchs W., Navon R., Kaback M. M., Kresse H. Tay-Sachs disease: one-step assay of beta-N-acetylhexosaminidase in serum with a sulphated chromogenic substrate. Clin Chim Acta. 1983 Oct 14;133(3):253–261. doi: 10.1016/0009-8981(83)90269-3. [DOI] [PubMed] [Google Scholar]
  8. Kaback M. M., Bailin G., Hirsch P., Roy C. Automated thermal fractionation of serum hexosaminidase: effects of alteration in reaction variables and implications for Tay-Sachs disease heterozygote screening. Prog Clin Biol Res. 1977;18:197–212. [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Lowden J. A., Zuker S., Wilensky A. J., Skomorowski M. A. Screening for carriers of Tay-Sachs disease: A community project. Can Med Assoc J. 1974 Aug 3;111(3):229–233. [PMC free article] [PubMed] [Google Scholar]
  11. Myerowitz R., Costigan F. C. The major defect in Ashkenazi Jews with Tay-Sachs disease is an insertion in the gene for the alpha-chain of beta-hexosaminidase. J Biol Chem. 1988 Dec 15;263(35):18587–18589. [PubMed] [Google Scholar]
  12. O'Brien J. S., Okada S., Chen A., Fillerup D. L. Tay-sachs disease. Detection of heterozygotes and homozygotes by serum hexosaminidase assay. N Engl J Med. 1970 Jul 2;283(1):15–20. doi: 10.1056/NEJM197007022830104. [DOI] [PubMed] [Google Scholar]
  13. Stirling J. L. Separation and characterisation of N-acetyl- -glucosaminidases A and P from maternal serum. Biochim Biophys Acta. 1972 Jun 22;271(1):154–162. doi: 10.1016/0005-2795(72)90143-2. [DOI] [PubMed] [Google Scholar]

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