title: Sequencing analysis of insulin receptor defects and detection of two novel mutations inINSRgene.
abstract: Mutations in the insulin receptor gene cause the inherited insulin resistant syndromes Leprechaunism and Rabson-Mendenhall syndrome. These recessive conditions are characterized by intrauterine and post-natal growth restrictions, dysmorphic features, altered glucose homeostasis, and early demise. The insulin receptor gene (INSR) maps to the short arm of chromosome 19 and is composed of 22 exons. Here we optimize the conditions for sequencing this gene and report novel mutations in patients with severe insulin resistance.
  
  
  METHODS
  PCR amplification of the 22 coding exons of theINSRgene was performed using M13-tailed primers. Bidirectional DNA sequencing was performed with BigDye Terminator chemistry and M13 primers and the product was analyzed on the ABI 3100 genetic analyzer. Data analysis was performed using Mutation Surveyor software comparing the sequence to a referenceINSRsequence (Genbank NC_000019).
  
  
  RESULTS
  We sequenced four patients with Leprechaunism or Rabson-Mendenhall syndromes as well as seven samples from normal individuals and confirmed previously identified mutations in the affected patients. Three of the four mutations identified in this group caused premature insertion of a stop codon. In addition, theINSRgene was sequenced in 14 clinical samples from patients with suspected insulin resistance and one novel mutation was found in an infant with a suspected diagnosis of Leprechaunism.
  
  
  CONCLUSIONS
  Leprechaunism and Rabson-Mendenhall syndrome are very rare and difficult to diagnose. Diagnosis is currently based mostly on clinical criteria. Clinical availability of DNA sequencing can provide an objective way of confirming or excluding the diagnosis.
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, USA; Division of Medical Genetics, Department of Pediatrics, University of Utah, Salt Lake City, UT, USA; Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA.
authors.forename: O
authors.initials: O
authors.lastname: Ardon
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, USA.
authors.forename: M
authors.initials: M
authors.lastname: Procter
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, USA.
authors.forename: T
authors.initials: T
authors.lastname: Tvrdik
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, USA; Division of Medical Genetics, Department of Pediatrics, University of Utah, Salt Lake City, UT, USA; Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA.
authors.forename: N
authors.initials: N
authors.lastname: Longo
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, USA; Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA.
authors.forename: R
authors.initials: R
authors.lastname: Mao
keywords: Donohue syndrome; Insulin receptor; Insulin resistance; Leprechaunism; Rabson–Mendenhall syndrome; Sequencing