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Molecular analysis of the CYP21A2 gene in Chinese patients with steroid 21-hydroxylase deficiency.
Clin Biochem. 2014 Apr; 47(6):455-63.CB

Abstract

OBJECTIVE

21-Hydroxylase deficiency (21-OHD) is the most common cause of congenital adrenal hyperplasia (CAH), a family of autosomal recessive disorders involving impaired cortisol synthesis. This study aimed to design a reliable and rational approach for identifying mutations in the CYP21A2 gene and to characterize the molecular basis of 21-OHD in 30 Chinese patients.

DESIGN AND METHODS

Copy number variations were investigated by multiplex ligation-dependent probe amplification (MLPA). Locus-specific polymerase chain reaction (PCR)/restriction endonuclease analysis was then used to verify CYP21A2 rearrangement products and prevent allele dropout. Direct sequencing of rearrangement products was performed to further refine recombination breakpoint locations. Direct sequencing of the entire CYP21A2 gene was used to detect microconversions.

RESULTS

We successfully characterized 60 CYP21A2 alleles from 30 patients with genetic defects. The most common one was intron 2 splice mutation (38.3%). Eighteen alleles with large gene deletions/conversions were identified, which accounted for nearly one-third (30.0%) of the genetic defects. Among these, three types of CYP21A1P/CYP21A2 chimeric genes (CH-1, CH-2, and CH-4) were characterized. Two novel CYP21A2 rearrangement genes were revealed and further demonstrated to be located downstream of the TNXB gene.

CONCLUSIONS

Our results indicate that the stepwise diagnostic procedure involving MLPA analysis, locus-specific PCR/restriction endonuclease analysis, and direct DNA sequencing can provide detailed genetic information about Chinese 21-OHD patients, which is helpful for characterizing structural rearrangements of CYP21A2.

Authors+Show Affiliations

State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China. Electronic address: jiangzhang784@163.com.State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, China. Electronic address: njxzf@126.com.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24503005

Citation

Ma, Dingyuan, et al. "Molecular Analysis of the CYP21A2 Gene in Chinese Patients With Steroid 21-hydroxylase Deficiency." Clinical Biochemistry, vol. 47, no. 6, 2014, pp. 455-63.
Ma D, Chen Y, Sun Y, et al. Molecular analysis of the CYP21A2 gene in Chinese patients with steroid 21-hydroxylase deficiency. Clin Biochem. 2014;47(6):455-63.
Ma, D., Chen, Y., Sun, Y., Yang, B., Cheng, J., Huang, M., Zhang, J., Zhang, J., Hu, P., Lin, Y., Jiang, T., & Xu, Z. (2014). Molecular analysis of the CYP21A2 gene in Chinese patients with steroid 21-hydroxylase deficiency. Clinical Biochemistry, 47(6), 455-63. https://doi.org/10.1016/j.clinbiochem.2014.01.019
Ma D, et al. Molecular Analysis of the CYP21A2 Gene in Chinese Patients With Steroid 21-hydroxylase Deficiency. Clin Biochem. 2014;47(6):455-63. PubMed PMID: 24503005.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular analysis of the CYP21A2 gene in Chinese patients with steroid 21-hydroxylase deficiency. AU - Ma,Dingyuan, AU - Chen,Yulin, AU - Sun,Yun, AU - Yang,Bing, AU - Cheng,Jian, AU - Huang,Meilian, AU - Zhang,Jin, AU - Zhang,Jingjing, AU - Hu,Ping, AU - Lin,Ying, AU - Jiang,Tao, AU - Xu,Zhengfeng, Y1 - 2014/02/03/ PY - 2013/05/18/received PY - 2014/01/16/revised PY - 2014/01/20/accepted PY - 2014/2/8/entrez PY - 2014/2/8/pubmed PY - 2016/4/1/medline KW - 21-Hydroxylase deficiency KW - CYP21A2 KW - Congenital adrenal hyperplasia KW - Mutation SP - 455 EP - 63 JF - Clinical biochemistry JO - Clin Biochem VL - 47 IS - 6 N2 - OBJECTIVE: 21-Hydroxylase deficiency (21-OHD) is the most common cause of congenital adrenal hyperplasia (CAH), a family of autosomal recessive disorders involving impaired cortisol synthesis. This study aimed to design a reliable and rational approach for identifying mutations in the CYP21A2 gene and to characterize the molecular basis of 21-OHD in 30 Chinese patients. DESIGN AND METHODS: Copy number variations were investigated by multiplex ligation-dependent probe amplification (MLPA). Locus-specific polymerase chain reaction (PCR)/restriction endonuclease analysis was then used to verify CYP21A2 rearrangement products and prevent allele dropout. Direct sequencing of rearrangement products was performed to further refine recombination breakpoint locations. Direct sequencing of the entire CYP21A2 gene was used to detect microconversions. RESULTS: We successfully characterized 60 CYP21A2 alleles from 30 patients with genetic defects. The most common one was intron 2 splice mutation (38.3%). Eighteen alleles with large gene deletions/conversions were identified, which accounted for nearly one-third (30.0%) of the genetic defects. Among these, three types of CYP21A1P/CYP21A2 chimeric genes (CH-1, CH-2, and CH-4) were characterized. Two novel CYP21A2 rearrangement genes were revealed and further demonstrated to be located downstream of the TNXB gene. CONCLUSIONS: Our results indicate that the stepwise diagnostic procedure involving MLPA analysis, locus-specific PCR/restriction endonuclease analysis, and direct DNA sequencing can provide detailed genetic information about Chinese 21-OHD patients, which is helpful for characterizing structural rearrangements of CYP21A2. SN - 1873-2933 UR - https://www.unboundmedicine.com/medline/citation/24503005/Molecular_analysis_of_the_CYP21A2_gene_in_Chinese_patients_with_steroid_21_hydroxylase_deficiency_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0009-9120(14)00047-2 DB - PRIME DP - Unbound Medicine ER -