1. Multiplex ligation-dependent probe amplification (MLPA) assay for the detection of CYP21A2 gene deletions/duplications in congenital adrenal hyperplasia: first technical report
- Author
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Ettore Capoluongo, Enrica Mello, Paola Concolino, Vincenzo Toscano, Cecilia Zuppi, Franco Ameglio, Concolino, P, Mello, E, Toscano, V, Ameglio, F, Zuppi, C, and Capoluongo, E.
- Subjects
Male ,Parents ,congenital, hereditary, and neonatal diseases and abnormalities ,endocrine system diseases ,Genotype ,Pseudogene ,Clinical Biochemistry ,Gene Dosage ,Prenatal diagnosis ,Biology ,urologic and male genital diseases ,Biochemistry ,Gene dosage ,Settore BIO/12 - BIOCHIMICA CLINICA E BIOLOGIA MOLECOLARE CLINICA ,Fetus ,Gene Duplication ,Gene duplication ,medicine ,CYP 21A2 ,Humans ,Multiplex ,Congenital adrenal hyperplasia ,Multiplex ligation-dependent probe amplification ,Alleles ,Southern blot ,Genetics ,Adrenal Hyperplasia, Congenital ,Siblings ,Biochemistry (medical) ,nutritional and metabolic diseases ,General Medicine ,Exons ,medicine.disease ,Molecular biology ,female genital diseases and pregnancy complications ,MLPA ,Mutation ,Female ,Steroid 21-Hydroxylase ,Nucleic Acid Amplification Techniques ,Gene Deletion - Abstract
Background More than 90% of the cases of Congenital Adrenal Hyperplasia (CAH) are associated with mutations in 21-hydroxylase gene ( CYP21A2 ). Up to now, large CYP21A2 rearrangements have been mainly detected by Southern blot analysis, although more rapid methods have been alternatively proposed. In this paper, we report the use of a multiplex ligation-dependent probe amplification (MLPA) method for easy and rapid detection of deletions/duplications in the CYP21A2 gene. Methods We collected 18 CAH Italian patients previously analyzed by gene sequencing and Southern blot technique. In addition, a prenatal diagnosis study was performed. Results Of the 7 known subjects with CYP21A2 deletions and 2 with gene duplications previously characterized in our laboratory, all were successfully identified by the MLPA analysis. In the prenatal diagnosis study, the MLPA assay was able to identify the presence of a CYP21A2 gene duplication in the fetus, as well in other two family members. Conclusion MLPA analysis represents a simple, rapid and sensitive tool for the detection of CYP21A2/CYP21A1P deletions/duplications in CAH molecular diagnosis. Compared to Southern blot, MLPA may be considered a high throughput analysis, allowing the simultaneous study of several samples in the same experiment and the investigation of both gene ( CYP21A2 ) and pseudogene ( CYP21A1P ) in each patient.
- Published
- 2009