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Molecular analysis of the CYP21A2 gene in dried blood spot samples.
Medicina (B Aires). 2020; 80(3):197-202.M

Abstract

Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder due to a deficiency of enzymes involved in cortisol biosynthesis. In more than 90% of cases, CAH is secondary to deleterious mutations in the CYP21A2 gene leading to 21-hydroxilase deficiency (21OHD). The CYP21A2 gene is located on the short arm of chromosome 6 (6p21·3) and encodes the cytochrome P450C21 enzyme. Neonatal screening programs detect the classic forms of CAH-21OHD quantifying 17OH-progesterone in dried blood spots (DBS). This test is very sensitive, but it has a low specificity, requiring a second sample to confirm the result. In these cases, a second-tier test in the same sample may be useful. Our aim was to evaluate a DNA extraction method from DBS and assess the performance of such DNA in the molecular analysis of the CYP21A2 gene mutations. Twelve individuals, who presumably had CAH based on the initial neonatal screening results, were analyzed using DNA extracted from freshly collected blood on EDTA and DBS. The CYP21A2 gene was analyzed by automated sequencing of all exons and intron boundaries and MLPA analysis in DBS. Molecular analysis results from both extraction methods were compared. In this study, we show that DNA extracted from neonatal screening DBS is a useful tool to define CYP21A2 gene mutations in 21-OHD diagnostic confirmation for the newborn screening program and that its results are comparable to traditional genotyping.

Authors+Show Affiliations

Laboratorio de Pesquisa Neonatal, Laboratorio de Biología Molecular y Diagnóstico, Servicio de Endocrinología, Hospital de Pediatría Prof. Dr. Juan P. Garrahan, Buenos Aires, Argentina.Laboratorio de Pesquisa Neonatal, Laboratorio de Biología Molecular y Diagnóstico, Servicio de Endocrinología, Hospital de Pediatría Prof. Dr. Juan P. Garrahan, Buenos Aires, Argentina.Laboratorio de Pesquisa Neonatal, Laboratorio de Biología Molecular y Diagnóstico, Servicio de Endocrinología, Hospital de Pediatría Prof. Dr. Juan P. Garrahan, Buenos Aires, Argentina.Laboratorio de Pesquisa Neonatal, Laboratorio de Biología Molecular y Diagnóstico, Servicio de Endocrinología, Hospital de Pediatría Prof. Dr. Juan P. Garrahan, Buenos Aires, Argentina.Laboratorio de Pesquisa Neonatal, Laboratorio de Biología Molecular y Diagnóstico, Servicio de Endocrinología, Hospital de Pediatría Prof. Dr. Juan P. Garrahan, Buenos Aires, Argentina.Laboratorio de Pesquisa Neonatal, Laboratorio de Biología Molecular y Diagnóstico, Servicio de Endocrinología, Hospital de Pediatría Prof. Dr. Juan P. Garrahan, Buenos Aires, Argentina.Laboratorio de Pesquisa Neonatal, Laboratorio de Biología Molecular y Diagnóstico, Servicio de Endocrinología, Hospital de Pediatría Prof. Dr. Juan P. Garrahan, Buenos Aires, Argentina. E-mail: marinorox@yahoo.com.

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article

Language

eng

PubMed ID

32442933

Citation

Marino, Silvia, et al. "Molecular Analysis of the CYP21A2 Gene in Dried Blood Spot Samples." Medicina, vol. 80, no. 3, 2020, pp. 197-202.
Marino S, Perez Garrido N, Ramírez P, et al. Molecular analysis of the CYP21A2 gene in dried blood spot samples. Medicina (B Aires). 2020;80(3):197-202.
Marino, S., Perez Garrido, N., Ramírez, P., Pujana, M., Dratler, G., Belgorosky, A., & Marino, R. (2020). Molecular analysis of the CYP21A2 gene in dried blood spot samples. Medicina, 80(3), 197-202.
Marino S, et al. Molecular Analysis of the CYP21A2 Gene in Dried Blood Spot Samples. Medicina (B Aires). 2020;80(3):197-202. PubMed PMID: 32442933.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular analysis of the CYP21A2 gene in dried blood spot samples. AU - Marino,Silvia, AU - Perez Garrido,Natalia, AU - Ramírez,Pablo, AU - Pujana,Matías, AU - Dratler,Gustavo, AU - Belgorosky,Alicia, AU - Marino,Roxana, PY - 2020/5/23/entrez PY - 2020/5/23/pubmed PY - 2020/6/9/medline KW - CYP21A2 KW - congenital adrenal hyperplasia KW - dried blood spot testing SP - 197 EP - 202 JF - Medicina JO - Medicina (B Aires) VL - 80 IS - 3 N2 - Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder due to a deficiency of enzymes involved in cortisol biosynthesis. In more than 90% of cases, CAH is secondary to deleterious mutations in the CYP21A2 gene leading to 21-hydroxilase deficiency (21OHD). The CYP21A2 gene is located on the short arm of chromosome 6 (6p21·3) and encodes the cytochrome P450C21 enzyme. Neonatal screening programs detect the classic forms of CAH-21OHD quantifying 17OH-progesterone in dried blood spots (DBS). This test is very sensitive, but it has a low specificity, requiring a second sample to confirm the result. In these cases, a second-tier test in the same sample may be useful. Our aim was to evaluate a DNA extraction method from DBS and assess the performance of such DNA in the molecular analysis of the CYP21A2 gene mutations. Twelve individuals, who presumably had CAH based on the initial neonatal screening results, were analyzed using DNA extracted from freshly collected blood on EDTA and DBS. The CYP21A2 gene was analyzed by automated sequencing of all exons and intron boundaries and MLPA analysis in DBS. Molecular analysis results from both extraction methods were compared. In this study, we show that DNA extracted from neonatal screening DBS is a useful tool to define CYP21A2 gene mutations in 21-OHD diagnostic confirmation for the newborn screening program and that its results are comparable to traditional genotyping. SN - 1669-9106 UR - https://www.unboundmedicine.com/medline/citation/32442933/Molecular_analysis_of_the_CYP21A2_gene_in_dried_blood_spot_samples_ L2 - http://www.medicinabuenosaires.com/PMID/32442933.pdf DB - PRIME DP - Unbound Medicine ER -