title: [Optimization of polymerase chain reaction assay combined with capillary electrophoresis to detect the pre- and full mutation of FMR1 gene].
abstract: OBJECTIVE
  To develop an efficient, high resolution PCR assay suitable for detection of the (CGG)n repeats of the fragile X mental retardation 1 (FMR1) gene by optimizing the PCR system in combination with capillary electrophoresis.
  
  
  METHODS
  Three standard samples and twelve samples that were verified by Southern blot analysis including both male and female in the normal, pre- and full mutation range were used in this study to evaluate the enhanced PCR system. All amplicons were electrophoresed by agarose, polyacrylamide and capillary electrophoresis to compare the results.
  
  
  RESULTS
  The enhanced PCR assay developed in this study was able to detect a full mutation with (CGG)n being larger than 260 repeats in a male. An expanded pre-mutation allele with (CGG)n as large as 183 repeats in a female was also amplified. The capillary electrophoresis method used in this study was able to distinguish two alleles with 1 CGG repeat difference and the results were reproducible.
  
  
  CONCLUSIONS
  A high resolution PCR assay is developed, which is much more efficient than the general PCR systems. It is suitable for the clinical screening of FMR1 gene and will greatly reduce the number of Southern blot analysis needed in clinical application.
authors.affiliation: Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai Institute for Pediatric Research, Shanghai 200092, P. R. China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Zhou
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.forename: Bing
authors.initials: B
authors.lastname: Xiao
authors.forename: Xu
authors.initials: X
authors.lastname: Han
authors.forename: Fei
authors.initials: F
authors.lastname: Long
authors.forename: Wen-ting
authors.initials: WT
authors.lastname: Jiang
authors.forename: Xing
authors.initials: X
authors.lastname: Ji
authors.forename: Jiong
authors.initials: J
authors.lastname: Tao
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics*; D006801:Humans; D008297:Male; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D051860:Fragile X Messenger Ribonucleoprotein 1; C497683:FMR1 protein, human