Analysis of the Type II 3β-Hydroxysteroid Dehydrogenase Gene in Patients with Nonclassical Congenital Adrenal Hyperplasia, Premature Pubarche, and Hyperandrogenism. † 407

Abstract

Mild 3β-hydroxysteroid dehydrogenase deficiency (NC3βHSDD) may cause premature pubarche (PP) and adolescent hyperandrogenism (HA). To test our hypothesis, we analyzed the type II 3β-HSD gene for mutations in 7 female patients (3PP, 4HA) who fit diagnostic criteria for NC3βHSDD, including steroid hormone responses to ACTH stimulation. An additional 37 female patients (22PP, 15HA) were also studied. Polymerase chain reaction(PCR) oligonucleotide primers specific for the type II 3β-HSD gene were used for single strand conformational polymorphism (SSCP) analysis of the coding region of the gene. Of the 44 patients studied, 4 patients (Table) were heterozygous for the same conformer. Sequence analysis of this conformer showed 2 point mutations in the 3′-untranslated region (3′-UTR), C8815G and A8921G. Restriction digest of PCR amplified genomic DNA confirmed these base changes occurred on the same allele. A fifth patient had a different conformer due to heterozygosity for a missense mutation (A167V). Neither unique conformer was detected in 100 healthy controls, p <0.008 by Fischer Exact Test.