Detection of AGXT gene mutations by denaturing high-performance liquid chromatography for diagnosis of hyperoxaluria type 1

D. Pirulli, M. Giordano, M. Lessi, A. Spanò, D. Puzzer, S. Zezlina, M. Boniotto, S. Crovella, F. Florian, M. Marangella, P. Momigliano-Richiardi, S. Savoldi, A. Amoroso

Research output: Contribution to journalArticle

Abstract

Primary hyperoxaluria type 1 is an autosomal recessive disorder of glyoxylate metabolism, caused by a deficiency of alanine:glyoxylate aminotransferase, which is encoded by a single copy gene (AGXT). The aim of this research was to standardize denaturing high-performance liquid chromatography, a new, sensitive, relatively inexpensive, and automated technique, for the detection of AGXT mutation. Denaturing high-performance liquid chromatography was used to analyze in blind the AGXT gene in 20 unrelated Italian patients with primary hyperoxaluria type 1 previously studied by other standard methods (single-strand conformation polymorphism analysis and direct sequencing) and 50 controls. Denaturing high-performance liquid chromatography allowed us to identify 13 mutations and the polymorphism at position 154 in exon I of the AGXT gene. Hence the method is more sensitive and less time consuming than single-strand conformation polymorphism analysis for the detection of AGXT mutations, thus representing a useful and reliable tool for detecting the mutations responsible for primary hyperoxaluria type 1. The new technology could also be helpful in the search for healthy carriers of AGXT mutations amongst family members and their partners, and for screening of AGXT polymorphisms in patients with nephrolithiasis and healthy populations.

Original languageEnglish
Pages (from-to)99-104
Number of pages6
JournalClinical and Experimental Medicine
Volume1
Issue number2
Publication statusPublished - 2001

Keywords

  • AGXT
  • Denaturing high-performance liquid chromatography
  • Heteroduplex
  • Hyperoxaluria type 1

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Clinical Biochemistry

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    Pirulli, D., Giordano, M., Lessi, M., Spanò, A., Puzzer, D., Zezlina, S., Boniotto, M., Crovella, S., Florian, F., Marangella, M., Momigliano-Richiardi, P., Savoldi, S., & Amoroso, A. (2001). Detection of AGXT gene mutations by denaturing high-performance liquid chromatography for diagnosis of hyperoxaluria type 1. Clinical and Experimental Medicine, 1(2), 99-104.