A PCR based X-chromosome inactivation assay for carrier detection in X-linked immunodeficiencies using differential methylation of the androgen receptor gene

Immunodeficiency. 1995;5(3):187-92.

Abstract

Carrier detection in X-linked immunodeficiencies (X-SCID, WAS, XLA) relies on the demonstration of non-random X inactivation patterns in blood cell lineages. Only a limited number of cells are available after cell separation methods. PCR-based techniques are therefore necessary to analyze active and inactive X chromosomes. Amplifying a polymorphic CAG repeat in the first exon of the androgen receptor gene after selective digestion of the active X chromosome with a methylation-sensitive restriction enzyme allows to distinguish between the paternal and maternal alleles and to identify their methylation status. DNA from B-, T-lymphocytes and total peripheral leukocytes of normal males, females and obligate carriers of X-linked immunodeficiencies were analyzed. The results of this PCR-based X inactivation assay are concordant with the standard methylation studies at the DXS255 locus using Southern blotting. This PCR assay provides a rapid and informative (heterozygosity > 90%) method in carrier detection of X-linked immunodeficiencies and other X-linked disorders, which show non-random X inactivation in cell lineages from the affected tissues.

MeSH terms

  • DNA / analysis
  • Dosage Compensation, Genetic*
  • Female
  • Genetic Carrier Screening / methods*
  • Genetic Linkage / genetics*
  • Humans
  • Immunologic Deficiency Syndromes / genetics*
  • Male
  • Methylation
  • Polymerase Chain Reaction / methods*
  • Receptors, Androgen / genetics*
  • X Chromosome / genetics*

Substances

  • Receptors, Androgen
  • DNA