Concurrent Confirmation and Differential Diagnosis of Congenital Adrenal Hyperplasia from Dried Blood Spots: Application of a Second-Tier LC-MS/MS Assay in a Cross-Border Cooperation for Newborn Screening.

Abstract

BACKGROUND/AIMS:

Newborn screening for congenital adrenal hyperplasia (CAH) is generally performed using 17- hydroxyprogesterone dissociation-enhanced, lanthanide fluorescence immunoassay (DELFIA®). The primary screening results must be confirmed due to high false-positive rates; however, the need to obtain a separate specimen can hamper early recognition, differential diagnosis and treatment. We aimed to develop a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) method that allows both the confirmation and differential diagnosis of CAH using the same dried blood spot (DBS) as in primary screening.

METHODS:

An LC-MS/MS assay for cortisol, 21-deoxycortisol, 11-deoxycortisol, 4-androstenedione and 17-hydroxyprogesterone was developed, validated and applied to a total of 163 DBS samples tested positive in primary newborn screening in a cross-border cooperation.

RESULTS:

Excellent baseline resolution and reliable determination of all analytes were achieved in DBS samples following simple sample preparation without derivatization. Of a total of 163 DBS samples tested positive in primary screening, the 21-hydroxylase-deficient form of CAH was confirmed in 1 sample.

CONCLUSIONS:

The present LC-MS/MS assay was successfully applied as a second-tier test in a cross-border cooperation for newborn screening. The assay allows concurrent confirmation and differential diagnosis of CAH and can be performed on the same DBS samples as in primary screening, enabling early diagnosis and treatment.

• Authors: Péter Monostori, Pál Szabó, Otilia Marginean, Csaba Bereczki, Eszter Karg
• Journal: Hormone research in paediatrics (Horm Res Paediatr) Vol. 84 Issue 5 Pg. 311-8 ( 2015) ISSN: 1663-2826 [Electronic] Switzerland
• PMID: 26397944 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2015 S. Karger AG, Basel.

Chemical References

• Steroid 21-Hydroxylase

Topics

• Adrenal Hyperplasia, Congenital (blood, diagnosis, economics)
• Chromatography, High Pressure Liquid
• Dried Blood Spot Testing (economics, methods)
• False Positive Reactions
• Female
• Humans
• Hungary
• Infant, Newborn
• International Cooperation
• Male
• Mass Spectrometry
• Neonatal Screening (economics, methods)
• Quality Control
• Reference Standards
• Reproducibility of Results
• Romania
• Steroid 21-Hydroxylase (blood)