APPLICATION OF MULTIPLEX LIGATION-DEPENDENT PROBE AMPLIFICATION (MLPA) IN DETECTING DELETION/DUPLICATION OF CYP21A2 CAUSING CONGENITAL ADRENAL HYPERPLASIA DISEASE
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Abstract
Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder usually caused by a deficiency of the enzyme 21-hydroxylase. The deficiency of the enzyme 21-hydroxylase occurs due to mutations in the CYP21A2 gene. Approximately 20-25% of patients with CAH carry mutations in copy number alterations such as deletions, duplications, or inversions of the CYP21A2 gene. The study was carried out with the aim of evaluating the CYP21A2 gene deletions using MLPA technique. 30 patients were diagnosed with congenital adrenal hyperplasia with 21- hydroxylase deficiency were collected; DNA extracted from blood samples was carried out; MLPA technique was performed to identify deletion/ duplication genes. The results found that 6 patients (20%) with homozygous deletion and 6 patients (20%) with heterozygous deletion. Among homozygous deletion mutants, deletion from exon 1 to exon 3 accounts for the highest percentage (50%), second is deletion from exon 1 to exon 7 (33.3%) and the remaning is deletion from exon 4 to exon 7 (16.7%).
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References
2. Speiser PW, Arlt W, Auchus RJ, Baskin LS, Conway GS, Merke DP, et al. Congenital Adrenal Hyperplasia Due to Steroid 21-Hydroxylase Deficiency: An Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab. 2018 Sep 27;103(11):4043–88.
3. Concolino P, Costella A. Congenital Adrenal Hyperplasia (CAH) due to 21-Hydroxylase Deficiency: A Comprehensive Focus on 233 Pathogenic Variants of CYP21A2 Gene. Mol Diagn Ther. 2018 Jun;22(3):261–80.
4. Schouten JP, McElgunn CJ, Waaijer R, Zwijnenburg D, Diepvens F, Pals G. Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. Nucleic Acids Res. 2002 Jun 15;30(12):e57.
5. Concolino P, Mello E, Toscano V, Ameglio F, Zuppi C, Capoluongo E. Multiplex ligation-dependent probe amplification (MLPA) assay for the detection of CYP21A2 gene deletions/ duplications in congenital adrenal hyperplasia: first technical report. Clin Chim Acta. 2009 Apr;402(1–2):164–70.
6. Balraj P, Lim PG, Sidek H, Wu LL, Khoo ASB. Mutational characterization of congenital adrenal hyperplasia due to 21-hydroxylase deficiency in Malaysia. J Endocrinol Invest. 2013Jun;36(6):366–74.
7. Vrzalová Z, Hrubá Z, St’ahlová Hrabincová E, Pouchlá S, Votava F, Kolousková S, et al. Identification of CYP21A2 mutant alleles in Czech patients with 21-hydroxylase deficiency. Int J Mol Med. 2010 Oct;26(4):595–603.
8. New MI, Abraham M, Gonzalez B, Dumic M, Razzaghy-Azar M, Chitayat D, et al. Genotype-phenotype correlation in 1,507 families with congenital adrenal hyperplasia owing to 21-hydroxylase deficiency. Proc Natl Acad Sci U S A. 2013 Feb 12;110(7):2611–6.