title: The use of high resolution melting analysis to detect Fabry mutations in heterozygous females via dry bloodspots.
abstract: BACKGROUND
  As an X-linked genetic disorder, Fabry disease was first thought to affect males only, and females were generally considered to be asymptomatic carriers. However, recent research suggests that female carriers of Fabry disease may still develop vital organ damage causing severe morbidity and mortality. In the previous newborn screening, from 299,007 newborns, we identified a total of 20 different Fabry mutations and 121 newborns with Fabry mutations. However, we found that most female carriers are not detected by enzyme assays.
  
  
  METHODS
  A streamlined method for high resolution melting (HRM) analysis was designed to screen for GLA gene mutations using a same PCR and melting program. Primer sets were designed to cover the 7 exons and the Chinese common intronic mutation, IVS4+919G>A of GLA gene.
  
  
  RESULTS
  The HRM analysis was successful in identifying heterozygous and hemizygous patients with the 20 surveyed mutations. We were also successful in using this method to test dry blood spots of newborns afflicted with Fabry mutations without having to determine DNA concentration before PCR amplification.
  
  
  CONCLUSIONS
  The results of this study show that HRM could be a reliable and sensitive method for use in the rapid screening of females for GLA mutations.
authors: Department of Pediatrics, Taipei Veterans General Hospital, Taipei, Taiwan.
authors: Chang-Long
authors: CL
authors: Tai
authors: Mei-Ying
authors: MY
authors: Liu
authors: Hsiao-Chi
authors: HC
authors: Yu
authors: Chiang-Chuan
authors: CC
authors: Chiang
authors: Hung
authors: H
authors: Chiang
authors: Jeng-Hung
authors: JH
authors: Suen
authors: Shu-Min
authors: SM
authors: Kao
authors: Yu-Hsiu
authors: YH
authors: Huang
authors: Tina Jui-Ting
authors: TJ
authors: Wu
authors: Chia-Feng
authors: CF
authors: Yang
authors: Fang-Chih
authors: FC
authors: Tsai
authors: Ching-Yuang
authors: CY
authors: Lin
authors: Jan-Gowth
authors: JG
authors: Chang
authors: Hong-Duo
authors: HD
authors: Chen
authors: Dau-Ming
authors: DM
authors: Niu
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D059788:Dried Blood Spot Testing / Q000379:methods*; D005091:Exons / Q000235:genetics; D000795:Fabry Disease / Q000097:blood* / Q000175:diagnosis / Q000235:genetics*; D005260:Female; D006579:Heterozygote; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D009154:Mutation; D009691:Nucleic Acid Denaturation; D044366:Transition Temperature