[Technique of PCR-ACRS for the detection of CYP21 gene mutations].
OBJECTIVE
To establish a rapid method of detecting CYP21 gene mutations.


METHODS
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.


RESULTS
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.


CONCLUSIONS
Analysis of CYP21 gene point mutations using PCR-ACRS is relatively simple, accurate and feasible.
Liao
Xiang-yun
XY
Xinhua Hospital, Shanghai Second Medical University, Shanghai Institute for Pediatric Research, Shanghai, 200092 PR China. xhkj@public.sta.net.cn
Zhang
Ya-fen
YF
Gu
Xue-fan
XF
D000312:Adrenal Hyperplasia, Congenital / Q000201:enzymology / Q000235:genetics*; D002681:China; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004252:DNA Mutational Analysis / Q000379:methods; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013255:Steroid 21-Hydroxylase / Q000235:genetics* / Q000378:metabolism
D004247:DNA; D013255:Steroid 21-Hydroxylase; D015252:Deoxyribonucleases, Type II Site-Specific; C027046:TCGA-specific type II deoxyribonucleases