Molecular and biochemical approaches in the identification of heterozygotes for homocystinuria

Michael Y. Tsai, Uttam Garg, Nigel S. Key, Naomi Q. Hanson, Anne Suh, Kerry Schwichtenberg

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51 Scopus citations


We compared biochemical and molecular methods for the identification of heterozygous carriers of mutations in the cystathionine β-synthase (CBS) gene. Eleven relatives of seven unrelated patients with homocystinuria due to homozygous CBS deficiency and controls were studied with respect to total homocysteine concentrations before and after methionine loading. In addition, we determined CBS activity in cultured skin fibroblasts and tested for the presence of five known mutations by a PCR-based method in these seven patients, their relatives and controls. The results demonstrate that measurement of homocysteine after methionine loading and assay of CBS enzyme activity in cultured fibroblasts identify most but not all heterozygotes. There was significant correlation between homocysteine concentrations and CBS activities only after methionine loading (r = 0.12, 0.48, 0.48 and 0.50 at 0, 4, 6 and 8 h, respectively). Among the homozygous patients, molecular approaches identified five T833C and two G919A mutations out of 14 independent alleles, confirming the studies of others that these represent the two most prevalent mutations. In addition, we found that three of six heterozygotes with the T833C allele had post-methionine loading homocysteine levels which overlapped with controls and of the other three, one (as well as an obligate heterozygote who did not carry any of the five mutant alleles tested) had CBS activity comparable to that of controls. These findings demonstrate that genotyping is useful as an adjunctive method for the diagnosis of the heterozygous carrier state of CBS deficiency.

Original languageEnglish (US)
Pages (from-to)69-77
Number of pages9
Issue number1
StatePublished - Apr 26 1996
Externally publishedYes

Bibliographical note

Funding Information:
This study was supported in part by Grant MOl-RR00400 from the National Center for Re-searchS ourcest o the Clinical ResearchC enter of the University of Minnesota Hospital and Clinic and by grantsf rom the Minnesota Medical Foundation of the University of Minnesota and the University of Minnesota’s Undergraduate Re-searchO pportunitiesP rogram. We thank Jeffrey Boutain for his technicala ssistance.


  • Amplification refractory mutation system
  • Cystathionine β-synthase
  • Genetic variants
  • Homocystinuria
  • Hyperhomocyst(e)inemia


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