Capillary zone electrophoresis in polymer networks of polymerase chain reaction-amplified oligonucleotides: the case of congenital adrenal hyperplasia.

The use of capillary zone electrophoresis (CZE) in polymer networks for the analysis of an 8 bp (base pair) deletion in congenital adrenal hyperplasia was investigated. Separations were performed in Tris-borate-EDTA buffer (pH 8.3) containing 6% liquid linear polyacrylamide as a sieving dynamic matrix and 10 microM ethidium bromide for improving DNA fragment separation. Easy analysis and detection of the 127 and 135 bp amplified fragments was accomplished. The capillary column can be used for > 50 analyses before degradation and loss of resolution. The results are comparable to those obtained by gel-slab zone electrophoresis in a 12%T, 4%C polyacrylamide matrix. The sensitivity, by simple UV absorption at 254 nm, is similar to that obtained in gel slabs by dye intercalation staining.

Molecular characterization of 21-hydroxylase deficiency in 70 Italian families.

Seventy Italian families affected by 21-hydroxylase deficiency were studied in order to evaluate the distribution of mutations. The coding P450c21B gene, the highly homologous P450c21A pseudogene and the linked C4A, C4B and DRB genes, mapping within the major histocompatibility complex region, were studied by multiple restriction analysis and in vitro amplification. In the affected individuals, 21.4% of the chromosomes were found to carry either gene deletions or large and small gene conversions. Our findings, consistent with previous reports in other ethnic groups, provide further evidence for the genetic heterogeneity of the disease.