A Set of Assays for the Comprehensive Analysis of FMR1 Alleles in the Fragile X-Related Disorders

J Mol Diagn. 2016 Sep;18(5):762-774. doi: 10.1016/j.jmoldx.2016.06.001. Epub 2016 Aug 12.


The diagnosis and study of the fragile X-related disorders is complicated by the difficulty of amplifying the long CGG/CCG-repeat tracts that are responsible for disease pathology, the potential presence of AGG interruptions within the repeat tract that can ameliorate expansion risk, the occurrence of variable DNA methylation that modulates disease severity, and the high frequency of mosaicism for both repeat number and methylation status. These factors complicate patient risk assessment. In addition, the variability in these parameters that is seen when patient cells are grown in culture requires their frequent monitoring to ensure reproducible results in a research setting. Many existing assays have the limited ability to amplify long alleles, particularly in a mixture of different allele sizes. Others are better at this, but are too expensive for routine use in most laboratories or for newborn screening programs and use reagents that are proprietary. We describe herein a set of assays to routinely evaluate all of these important parameters in a time- and cost-effective way.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Alleles*
  • Cell Line
  • DNA Methylation
  • Female
  • Fragile X Mental Retardation Protein / genetics*
  • Fragile X Syndrome / diagnosis*
  • Fragile X Syndrome / genetics*
  • Genetic Testing* / methods
  • Humans
  • Male
  • Mutation*
  • Polymerase Chain Reaction / methods
  • Promoter Regions, Genetic
  • Trinucleotide Repeat Expansion
  • Trinucleotide Repeats
  • Workflow


  • Fragile X Mental Retardation Protein