Technique of PCR-ACRS for the detection of CYP21 gene mutations / 中华医学遗传学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To establish a rapid method of detecting CYP21 gene mutations.</p><p><b>METHODS</b>Fifty Chinese patients with 21-hydroxylase deficiency and some of their families were investigated. Blood samples were obtained for extraction of peripheral blood lymphocytes. A search for restriction sites discriminating between the morbid and the normal in CYP21 gene was made by the computer program DNAssist. PCR-based amplication-created restriction site(PCR-ACRS) was performed at I172N and R356W which are not natural recognition sequence. In addition, I172N and R356W were analysed in five families which conform to the applicability of PCR-ACRS.</p><p><b>RESULTS</b>In 50 identified 21-hydroxylase deficient Chinese patients, 21 were found to have I172 N (3 were homozygote, 18 were heterozygote); 8 were found to have R356W, all of them were heterozygote. By analysing the families, the findings were consistent with the characteristics of autosomal recessive genetic deficiency.</p><p><b>CONCLUSION</b>Analysis of CYP21 gene point mutations using PCR-ACRS is relatively simple, accurate and feasible.</p>