The Use of Recombinant DNA Techniques for the Diagnosis of Familial Hypercholesterolaemia

  • S. Humphries
  • R. Taylor
  • M. Jeenah
  • M. Seed


In the UK, about 5% of patients with familial hypercholesterolaemia have a detectable deletion or rearrangement of part of the LDL-receptor gene. This results in the detection of shorter or abnormal sized fragments of the LDL-receptor gene in a Southern blot hybridization. This can be used to follow the inheritance of the defective gene, and for diagnosis in the families of these individuals. In the families of the rest of the patients, diagnosis may be possible using linked restriction fragment length polymorphisms (RFLPs) detected with the LDL-receptor probe. There are now ten common RFLPs of the LDL-receptor gene, with variable sites in the 3′ half of the gene. Over 80% of patients are heterozygous for at least one of these RFLPs, and therefore potentially informative for DNA diagnosis. For a foetus at risk of homozygous familial hypercholesterolaemia, antenatal diagnosis may also be possible using these methods. However, family studies require samples to be available from affected or unaffected relatives of the patient, and this limits the applicability of the tests. For some mutations, the base pair change causing the defect in the LDL-receptor itself creates or destroys a site for a restriction enzyme. Such ‘mutation-specific’ RFLPs could be used for population screening, but so far have only been reported for the familial hypercholesterolaemia mutation that is common in Lebanon. In the future it may be possible to develop mutation-specific oligonucleotide probes for the diagnosis of familial hypercholesterolaemia. These would be appropriate for population screening or screening patients with hyperlipidaemia. This information may be useful if different mutations require different therapeutic strategies.


Familial Hypercholesterolaemia Restriction Fragment Length Polymorphism Defective Gene Lipid Research Clinic Program Pvuii Polymorphism 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Antonarakis, S. E., Kazazian, H. H. and Orkin, S. H. DNA polymorphism and molecular pathology of the human globin gene clusters. Hum. Genet. 69 (1985) 1–14PubMedCrossRefGoogle Scholar
  2. Armston, A. E., Iverson, S. A., Burke, J. F. Diagnosis of familial hypercholesterolaemia using DNA probes for the low-density lipoprotein (LDL) receptor gene. Ann. Clin. Biochem. 25 (1988) 142–149PubMedGoogle Scholar
  3. Berg, K., Pedersen, J. C., Borresen, A. I., Heiberg, A. and Solaas, M. H. Close linkage between a common DNA polymorphism of the low-density lipoprotein (LDL) receptor gene and its use in diagnosis. Cytogenet. Cell. Genet. 40 (1985) 581–582Google Scholar
  4. Botstein, D., White, R. L., Skolnick, M. and Davis, R. W. Construction of a genetic linkage map using restriction fragment length polymorphisms. Am. J. Hum. Genet. 32 (1980) 314–331PubMedGoogle Scholar
  5. Brink, P. A., Steyn, L. T., Bester, A. J. and Steyn, K. Linkage disequilibrium between a marker on the low-density lipoprotein receptor and high cholesterol levels. South Afr. Med. J. 70 (1986) 80–82Google Scholar
  6. Brink, P. A., Steyn, L. T., Coetzee, G. A. and Van der Westhuyzen, D. R. Familial Hypercholesterolaemia in South African Afrikaaners: PvuII and StuI DNA polymorphisms in the LDL-receptor gene consistent with a predominating founder gene effect. Hum. Genet. 77 (1987) 32–35PubMedCrossRefGoogle Scholar
  7. Conner, B. J., Reyes, A. A., Morin, C., Itakura, K., Teplitz, R. L. and Wallace, R. B. Detection of sickle cell βS globin allele by hybridisation with synthetic oligonucleotides. Proc. Natl. Acad. Sci. USA 80 (1983) 278–282PubMedCrossRefGoogle Scholar
  8. Funke, H., Klug, J., Frossard, P., Coleman, R. and Assmann, G. PstI RFLP close to the LDL receptor gene. Nucl. Acid Res. 14 (1986a) 7820CrossRefGoogle Scholar
  9. Funke, H., Rust, S. and Assmann, G. Detection of apolipoprotein E variants by an oligonucleotide “melting” procedure. Clin. Chem. 32 (1986b) 1285–1289PubMedGoogle Scholar
  10. Geisel, J., Weisshaar, B., Oette, K., Mechtel, M. and Doerfler, W. Double MspI RFLP in the human LDL-receptor gene. Nucl. Acid. Res. 15 (1987) 3943CrossRefGoogle Scholar
  11. Gluek, C. J., Heckmann, F., Schoenfeld, M., et al. Neonatal familial type II hyperlipoproteinaemia: cord blood cholesterol in 1800 births. Metabolism 20 (1971) 597–608CrossRefGoogle Scholar
  12. Hobbs, H. H., Lehrman, M. A., Yamamoto, T. and Russell, D. W. Polymorphism and evolution of Alu sequences in the human low density lipoprotein receptor gene. Proc. Natl. Acad. Sci. USA 82 (1985) 7651–7655PubMedCrossRefGoogle Scholar
  13. Hobbs, H. H., Esser, V. and Russell, D. W. AvaII polymorphism in the human LDL receptor gene. Nucl. Acid Res. 15 (1987a) 379CrossRefGoogle Scholar
  14. Hobbs, H. H., Brown, M. S., Russell, P. W., Davigon, J. and Goldstein, J. L. Deletion in LDL receptor gene occurs in majority of French Canadians with FH. N. Engl. J. Med. 317 (1987b) 734–737CrossRefGoogle Scholar
  15. Horsthemke, B., Kessling, A. M., Seed, M., Wynn, V., Williamson, R. and Humphries, S. E. Identification of a deletion in the low density lipoprotein (LDL) receptor gene in a patient with familial hypercholesterolaemia. Hum. Genet. 71 (1985) 75–78PubMedCrossRefGoogle Scholar
  16. Horsthemke, B., Beisiegel, U., Dunning, A., Williamson, R. and Humphries, S. Nonhomologous crossing-over between two alu-repetitive DNA sequences in the LDL-receptor gene: A possible mechanism for a novel mutation in a patient with familial hypercholesterolaemia. Eur. J. Biochem. 164 (1987a) 77–81CrossRefGoogle Scholar
  17. Horsthemke, B., Dunning, A. and Humphries, S. Identification of deletions in the human low-density lipoprotein (LDL) receptor. J. Med. Genet. 24 (1987b) 144–147CrossRefGoogle Scholar
  18. Humphries, S. E., Kessling, A. M., Horsthemke, B., Donald, J. A., Seed, M., Jowett, N., Holm, M., Galton, D. J., Wynn, V. and Williamson, R. A common DNA polymorphism of the low density lipoprotein (LDL) receptor gene and its use in diagnosis. Lancet 1 (1985) 1003–1005PubMedCrossRefGoogle Scholar
  19. Kotze, M. J., Langenhoven, E., Dietzsch, E., and Retief, A. E. An RFLP associated with the low-density lipoprotein receptor gene (LDLR). Nucl. Acid Res. 15 (1987) 37Google Scholar
  20. Kotze, M. J., Retief, A. E., Brink, P. A. and Weich, H. F. H. A DNA polymorphism in the human low-density lipoprotein receptor gene. S. Afr. J. Med. 70 (1986) 77–79Google Scholar
  21. Kwiterovich, P. O., Levy, R. I. and Frederickson, D. S. Diagnosis of familial type-II hyperlipoproteinaemia. Lancet 1 (1973) 118–121PubMedCrossRefGoogle Scholar
  22. Lehrman, M. A., Schneider, W. J., Sudhof, T. C., Brown, M. S., Goldstein, J. L. and Russell, D. W. Mutations in LDL-receptor: Alu-Alu recombination deletes exons encoding transmembrane and cytoplasmic domains. Science 227 (1985a) 140–146CrossRefGoogle Scholar
  23. Lehrman, M. A., Goldstein, J. L., Brown, M. S., Russell, D. W. and Schneider, W. J. Internalization-defective LDL-receptors produced by genes with nonsense and frameshift mutations that truncate the cytoplasmic domain. Cell 47 (1985b) 735–743CrossRefGoogle Scholar
  24. Lehrmann, M. A., Goldstein, J. L., Russell, D. W. and Brown, M. S. Duplication of seven exons in LDL receptor gene caused by Alu-Alu recombination in a subject with familial hypercholesterolemia. Cell 48 (1987a) 827–835CrossRefGoogle Scholar
  25. Lehrman, M. A., Schneider, W. J., Brown, M. S., Davis, C. G., Elhammer, A., Russell, D. W. and Goldstein, J. L. The Lebanese allele at the low-density lipoprotein receptor locus. Nonsense mutation produces truncated receptor that is retained in endoplasmic reticulum. Biol. Chem. 262 (1987b) 401–410Google Scholar
  26. Leitersdorf, E. and Hobbs, H. H. Human LDL receptor gene: two ApaLI RFLPs. Nucl. Acid Res. 15 (1987) 2982Google Scholar
  27. Leonard, J. V., Whitelaw, A. G. L., Wolff, O. H., Lloyd, J. K. and Slack, J. Diagnosing familial hypercholesterolemia in children by measuring serum cholesterol. Br. Med. J. (1977) 1566–1568Google Scholar
  28. Leppert, M. F., Hasstedt, S. J., Holm, T., O’Connell, P., Wu, L., Ash, O., Williams, R. R. and White, R. A DNA probe for the LDL receptor gene is tightly linked to hypercholesterolaemia in a pedigree with early coronary disease. Am. J. Hum. Genet. 39 (1986) 300–306PubMedGoogle Scholar
  29. Lipid Research Clinics Program. The Lipid Research Clinics Coronary Primary Prevention Trial Results. II. The relationship of reduction in incidence of coronary heart disease to cholesterol lowering. J. Am. Med. Assoc. 251 (1984) 356–374Google Scholar
  30. Morris, S. W. and Price, W. H. DNA sequence polymorphisms in the apolipoprotein A-I/ C-III gene cluster. Lancet 2 (1985) 1127–1128PubMedCrossRefGoogle Scholar
  31. Old, J. M., Ward, R. H. T., Karagozlu, F., Petrou, M., Modell, B. and Weatherall, D. J. First-trimester fetal diagnosis for haemoglobinopathies: three cases. Lancet 2 (1982) 1413–1416PubMedCrossRefGoogle Scholar
  32. Steyn, L. T., Pretorius, A., Brink, P. A. and Bester, A. J. RFLP for the human LDL receptor gene (LDLR): BstEII. Nucl. Acid. Res. 15 (1987) 4702CrossRefGoogle Scholar
  33. Yamamoto, T., Davis, L. G., Brown, M. S., Schneider, W. J., Casey, M. L., Goldstein, J. L. and Russell, D. W. The human LDL-receptor: a cysteine-rich protein with multiple Alu sequences in its mRNA. Cell 39 (1984) 27–38PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 1988

Authors and Affiliations

  • S. Humphries
    • 1
  • R. Taylor
    • 1
  • M. Jeenah
    • 1
  • M. Seed
    • 2
  1. 1.Charing Cross Sunley Research CentreLondonUK
  2. 2.Charing Cross Hospital Medical SchoolLondonUK

Personalised recommendations