ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)
"Biotechnologia Acta" v. 7, no 1, 2014
https://doi.org/10.15407/biotech7.01.075
Р. 75-79, Bibliography 11, Ukrainian
Universal Decimal classification: 606:61, 602
ANALYSIS CYP21A2 GENE MUTATIONS TECHNIQUE IN PATIENTS WITH CONGENITAL ADRENAL HYPERPLASIA
Chernushyn S. Yu., Livshits L. A.
Institute of Molecular Biology and Genetics of National Academy of Sciences of Ukraine, Kyiv, Ukraine
The technique of CYP21A2 gene mutation analysis, which can be applicable for pre- and postnatal diagnosis of congenital adrenal hyperplasia various types was developed.
The analysis of extended CYP21A2 gene deletions and mutations 655A/C>G, 999T>A, 1994C>T, 2108C>T, which lead to congenital adrenal hyperplasia simple virilysing and salt wasting forms, and mutations 89C> T and 1683G> T, which lead to non-classical forms of the illness was proposed. This technique is based on analysis of the allele specific polymerase chain reaction and restriction fragment length polymorphism.
The technique has passed quality control on available DNA samples. Thus this may be suggested for DNA diagnostics (including prenatal) in high-risk families and for analysis of heterozygous carriers illness. Using the developed technique nine alleles with CYP21A2 gene deletion, 4 alleles with 655A/C>G mutation and 1 allele with 2108C> T mutation were detected in seven patients and their parents.
Key words: congenital adrenal hyperplasia, enzyme 21-hydroxylase, CYP21A2 gene.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2014
References
1. Merke D. P., Bornstein S. R. Congenital adrenal hyperplasia. Lancet. 2005, V. 365, 2125–2136.
2. Levine L. S. Congenital adrenal hyperplasia. Pediatr. Rev. 2000, V. 21, Р. 159–170.
3. Ingelman-Sundberg M., Daly A.K., Nebert D.W., Sim S.C. The Human Cytochrome P450 (CYP) Allele Nomenclature Database. Available at http://www.cypalleles.ki.se/
4. Baumgartner-Parzer S. M., Nowotny P., Heinze G. Waldh?usl W, Vierhapper H. Carrier frequency of congenital adrenal hyperplasia (21-hydroxylase deficiency) in a middle European population. J. Clin. Endocrinol. Metab. 2005, V. 90, Р. 775–778.
5. New M. I. Extensive clinical experience: nonclassical 21-hydroxylase deficiency. J. Clin. Endocrinol. Metab. 2006, V. 91, Р. 4205–4214.
6. Speiser P. W., White P. Congenital adrenal hyperplasia, New Eng. J. Med. 2003, V. 349, Р. 776–788.
7. Charmandari E., Eisenhofer G., Mehlinger S. L., Carlson A., Wesley R., Keil M. F., Chrousos G. P., New M. I., Merke D. P. Adrenomedullary function may predict phenotype and genotype in classic 21-hydroxylase deficiency. J. Clin. Endocr. Metab. 2002, V. 87, Р. 3031–3037.
8. Haider S., Islam B., D’Atri V., Sgobba M., Poojari C., Sun L., Yuen T., Zaidi M., New M. I. Structure-phenotype correlations of human CYP21A2 mutations in congenital adrenal hyperplasia. Proc. Natl. Acad. Sci. 2013, V. 110, Р. 2605–2610.
9. Maniatis Т., Fritsch E F., Sambrook J. Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory. 1982, Р.265 .
10. Vasudevamurthy M. K., Lever M., George P. M., Morison K. R. Betaine structure and the presence of hydroxyl groups alters the effects on DNA melting temperatures. Biopolymers. 2009, V. 91, 85–94.
11. Stikkelbroeck N. M., Hoefsloot L. H., de Wijs I. J., Otten B. J., Hermus A. R., Sistermans E. A. CYP21 Gene Mutation Analysis in 198 Patients with 21-Hydroxylase Deficiency in The Netherlands: Six Novel Mutations and a Specific Cluster of Four Mutations. J. Clin. Endocrinol. Metabol. 2003, V. 88, Р. 83852–3859.