摘要
目的建立一种等位基因特异PCR结合测序检测CYP21A2基因突变的方法。方法通过对CYP21A2基因和相应假基因CYP21AP序列进行同源性比较，同时设计等位基因特异PCR引物与可扩增CYP21A2和CYP21A2P基因的同源性引物。选择4例先天性肾上腺皮质增生症患者和5名正常对照，比较等位基因特异PCR引物与同源引物测序结果。结果等位基因特异PCR引物扩增和测序分析结果显示，4例患者均存在突变，分别为IVS2—13A／C〉G、Arg356Trp和Arg149Pro，对照序列正常。同源引物扩增分析只能明确Arg149Pro突变，患者和正常对照均存在IVS2—13A／C〉G和Arg356Trp突变。结论等位基因特异PCR技术结合测序可简便可靠地进行CYP21A2基因突变检测，具有临床应用推广价值。
Objective To establish an allele-specific PCR method for detect screening of CYP21A2 gene mutation. Methods Allele-specific PCR primers and analogy primers were designed based on the sequence alignment of CYP21A2 and CYP21AP genes. Genomic DNA was extracted from blood specimens of 4 patients with 21-hydroxylase deficiency and 5 healthy controls and respectively amplified with allele- specific PCR primers and analogy primers and sequenced. Results Mutations of CYP21A2 including IVS2—13A/C〉G, Arg356Trp and Arg149Pro were found with the established method in all of the 4 patients but not in the healthy controls. When detected with the analogy primers set, IVS2- 13A/C〉G and Arg356Trp were observed in both patients and healthy controls. Conclusion The allele-specific PCR-based method is a simple, effective and reliable method for the detection of CYP21A2 gene mutation.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2014年第4期479-482,共4页
Chinese Journal of Medical Genetics
