Received on September 3, 2004
Accepted on November 22, 2004

Molecular Diagnostics and Genetics

Rapid Second-Tier Molecular Genetic Analysis for Congenital Adrenal Hyperplasia attributable to Steroid 21-Hydroxylase Deficiency

¹ Labor Becker, Olgemöller und Kollegen, Munich, Germany
² Klinik mit Poliklinik für Kinder und Jugendliche, Friedrich-Alexander-Universität, Erlangen, Germany
³ Dr. von Haunersches Kinderspital, Ludwig-Maximilians-Universität, Munich, Germany

^* To whom correspondence should be addressed. E-mail: b.olgemoeller{at}labor-bo.de.

Background: Neonatal screening for steroid 21-hydroxylase (CYP21) deficiency is performed to identify congenital adrenal hyperplasia (CAH). The immunologic assay for 17-hydroxyprogesterone (17-OHP) has a high rate of false positives. We assessed the potential for increasing the specificity for CAH by use of a second step involving analysis of the CYP21 gene.

Methods: Between January 1999 and December 2003, a total of 810 000 newborns were screened. Of these, 7920 had to be retested because their 17-OHP values were above the cutoff of the assay. Sixty-one had positive 17-OHP values in their recall samples and were diagnosed as having CAH. We used a rapid assay for common mutations of the CYP21 gene to analyze these 61 samples. In a prospective study, 198 consecutive samples that had increased 17-OHP and 100 samples that had normal 17-OHP concentrations were genotyped.

Results: Fifty-nine of 61 cases diagnosed as having CAH were confirmed genetically as CYP21 deficiencies. One patient had a 3-hydroxysteroid-dehydrogenase deficiency, and one patient carried no CYP21 mutations. The 198 increased 17-OHP results were designated as false positives after immunologic testing of recall samples. None of these samples exhibited the genetic pattern consistent with CYP21 deficiency.

Conclusions: If samples with increased 17-OHP values were screened genetically, the number of retests would decrease by 90%, but the overall sensitivity of CAH screening would remain the same. Adding a second-tier genetic step would require a modest increase in costs, but is counterbalanced by fewer recalls, less clinical follow-up, and a reduction in unnecessary worry for families.