Briefly, we described 2 sisters in a family representing manifestations of Wiskott-Aldrich syndrome (WAS). The older sister had suffered from recurrent infections, small-sized thrombocytopenia, petechiase and purpura, and eczema for 7 years. The younger sister had the same manifestations, and died of intracranial bleeding at age 2 years. All the laboratory data on the 2 patients and the result of the sialophorin analysis were compatible with WAS, although they were females. Polymerase chain reaction (PCR) analysis of the sialophorin gene and singlestrand conformation polymorphism (SSCP) analysis of the PCR product demonstrated that there were no detectable size-changes or electrophoretic mobility changes in the DNA from both parents. Studies on the mother-daughter transmission of the X chromosome using a pERT84-MaeIII polymorphic marker mapped at Xp21, and HPRT gene polymorphism at Xq26, suggested that each sister had inherited a different X chromosome from the mother.
We subsequently tried reverse-transcriptase (RT)-PCR analysis against the WASP gene of our patient's peripheral lymphocyte to elucidate whether the older sister had any mutations in the WASP gene. We used two sets of PCR primers which overlay the coding region of the WASP cDNA . The sequences and locations of the primers are WASP1F: 5Π-CAGAGAAGACAAGGGCAGAA-3Π, nucleotide position 9-28; WASP1R: 5Π-TAAGTTTAGAGGTCTCGGCG-3Π, nucleotide position 883-902; WASP2F: 5Π-AGATCT-GCGGAGTCTGTTCT-3Π, nucleotide position 829-848; and WASP2R: 5Π-ACAGGGCAGCAAGTAACTCA-3Π, nucleotide position 1546-1565. As shown in Figure , there is no detectable size-change.
Next, we performed parental-origin analysis of chromosome 16 of DNAs in this family to make clear whether the aberration of the sialophorin gene, including the noncoding region, caused our patients' disorder. We used seven set of primers, i.e., D16S402, D16S403, D16S404, D16S406, D16S407, D16S420, and the sialophorin CA repeat, located on chromosome 16.