High resolution melting for the identification of mutations in the iron responsive element of the ferritin light chain gene

Emanuela Castiglioni, Nadia Soriani, Domenico Girelli, Clara Camaschella, Ivana Spiga, Matteo G. Della Porta, Maurizio Ferrari, Laura Cremonesi

Research output: Contribution to journalArticle

Abstract

Background: Among the causes of hyperferritinemia, hereditary hyperferritinemia cataract syndrome (HHCS) is an autosomal dominant disease characterized by distinctive cataracts and high serum ferritin. It is caused by mutations in the iron responsive element (IRE) of the ferritin light chain gene (FTL). Methods: To speed up and simplify mutational scanning in this genomic region, we developed a protocol based on high-resolution melting (HRM) analysis. Results: Validation was carried out using 18 wild-type and 14 DNA samples carrying different mutations, each analyzed in replicates of 20. The method allowed for correct identification and genotyping of all mutant samples, and each variant generated a specific profile distinguishable from the wild type. A 5.5% proportion of false positive results were obtained. In addition, in two patients with HHCS, two new mutations were identified by HRM based on an altered melting profile. These mutations were subsequently characterized by direct sequencing (7C>G+40A>G and 49A>C). Conclusions: The high reliability of HRM in detecting known and new DNA variations indicate that this could be an effective and sensitive method for molecular scanning of mutations in the IRE of the FTL gene in patients presenting with either HHCS or unexplained hyperferritinemia.

Original languageEnglish
Pages (from-to)1415-1418
Number of pages4
JournalClinical Chemistry and Laboratory Medicine
Volume48
Issue number10
DOIs
Publication statusPublished - Oct 1 2010

Keywords

  • ferritin light chain gene
  • hereditary hyperferritinemia cataract syndrome
  • high resolution melting
  • iron responsive element

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

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