A fluorescent method for detecting low-grade 11patUPD mosaicism in Beckwith-Wiedemann syndrome

Mol Cell Probes. 2003 Dec;17(6):295-9. doi: 10.1016/j.mcp.2003.07.002.

Abstract

The quantitative evaluation of mosaicism for uniparental disomy (UPD) involving a restricted chromosomal region requires the availability of a sensitive and reproducible method that is capable of detecting even a small percentage of disomic cells and avoiding false positive and false negative results. The occurrence of UPD is usually monitored by means of the parent-proband segregation analysis of microsatellites mapping to the target region. We here describe the quantitative blood cell evaluation of segmental mosaic UPD11, a marker of Beckwith-Wiedemann syndrome, by means of the segregation analysis of 11p15 microsatellites using both radioactive and fluorescence-based techniques. As the greater amplification efficiency of the shorter allele in heterozygous subjects may bias the correct evaluation of disomy, the mean short/long allele ratio was established at three loci of each of 30 normal heterozygous subjects, as well as the peak As/Al area in the presence of 50% of each allele. The interval was defined using a 5% level of significance. The results show that the fluorescence-based technique is superior to radioactivity in detecting the subtle allelic imbalances present in low-grade mosaicism conditions.

MeSH terms

  • Alleles
  • Beckwith-Wiedemann Syndrome / genetics*
  • Chromosomes, Human, Pair 11
  • Fluorescent Dyes
  • Genomic Imprinting / genetics
  • Humans
  • Isotope Labeling
  • Microsatellite Repeats / genetics*
  • Mosaicism / genetics*
  • Uniparental Disomy / genetics*

Substances

  • Fluorescent Dyes