Analysis of the chimeric CYP21P/CYP21 gene in steroid 21-hydroxylase deficiency

Hsien Hsiung Lee, Jan Gowth Chang, Chang Hai Tsai, Fuu Jen Tsai, Hsiang Tai Chao, Bon Chu Chung

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Background: A single nonfunctional chimeric gene with its 5' and 3' ends corresponding to CYP21P and CYP21, respectively, is caused by unequal gene crossover in the CYP21 genes during meiosis. The presence of the chimeric CYP21P/CYP21 molecule can not be detected by conventional PCR methods and therefore may be lost in PCR amplification. This leads to a false result and diagnostic discordance. Methods: We developed a rapid and direct method to detect a chimeric CYP21P/CYP21 gene that uses a 3'-specific primer for the CYP21 gene and two different 5' primers for both CYP21 and CYP21P to amplify the wild-type CYP21 and the chimeric CYP21P/CYP21 genes. A secondary PCR that can differentiate the chimeric from the wild-type gene was also performed. The PCR product was directly analyzed on agarose gel. Results: After careful titration, we found that earlier failure to detect the chimeric CYP21P/CYP21 gene could be caused by unequal concentrations of two independent alleles as the PCR template or by the lack of primers to amplify chimeric molecules. We successfully amplified the chimeric gene using our improved method. Conclusions: The chimeric CYP21P/CYP21 is present in a large portion of congenital adrenal hyperplasia patients. By adding a CYP21P/CYP21-specific primer, we were able to amplify and detect both homozygous and heterozygous chimeric genes. Therefore, our new PCR-based assay is a more effective way to analyze congenital adrenal hyperplasia mutations. (C) 2000 American Association for Clinical Chemistry.

Original languageEnglish
Pages (from-to)606-611
Number of pages6
JournalClinical Chemistry
Volume46
Issue number5
Publication statusPublished - Jun 6 2000

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Steroid 21-Hydroxylase
Genes
Polymerase Chain Reaction
Congenital Adrenal Hyperplasia
Congenital adrenal hyperplasia due to 21 hydroxylase deficiency
Molecules
Meiosis
Titration
Sepharose
Amplification
Assays
Gels
Alleles

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Lee, H. H., Chang, J. G., Tsai, C. H., Tsai, F. J., Chao, H. T., & Chung, B. C. (2000). Analysis of the chimeric CYP21P/CYP21 gene in steroid 21-hydroxylase deficiency. Clinical Chemistry, 46(5), 606-611.

Analysis of the chimeric CYP21P/CYP21 gene in steroid 21-hydroxylase deficiency. / Lee, Hsien Hsiung; Chang, Jan Gowth; Tsai, Chang Hai; Tsai, Fuu Jen; Chao, Hsiang Tai; Chung, Bon Chu.

In: Clinical Chemistry, Vol. 46, No. 5, 06.06.2000, p. 606-611.

Research output: Contribution to journalArticle

Lee, HH, Chang, JG, Tsai, CH, Tsai, FJ, Chao, HT & Chung, BC 2000, 'Analysis of the chimeric CYP21P/CYP21 gene in steroid 21-hydroxylase deficiency', Clinical Chemistry, vol. 46, no. 5, pp. 606-611.
Lee HH, Chang JG, Tsai CH, Tsai FJ, Chao HT, Chung BC. Analysis of the chimeric CYP21P/CYP21 gene in steroid 21-hydroxylase deficiency. Clinical Chemistry. 2000 Jun 6;46(5):606-611.
Lee, Hsien Hsiung ; Chang, Jan Gowth ; Tsai, Chang Hai ; Tsai, Fuu Jen ; Chao, Hsiang Tai ; Chung, Bon Chu. / Analysis of the chimeric CYP21P/CYP21 gene in steroid 21-hydroxylase deficiency. In: Clinical Chemistry. 2000 ; Vol. 46, No. 5. pp. 606-611.
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