CAJ | 학술논문

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.
목적 건립일충등위기인특이PCR결합측서검측CYP21A2기인돌변적방법.방법 통과대CYP21A2기인화상응가기인CYP21AP서렬진행동원성비교,동시설계등위기인특이PCR인물여가확증CYP21A2화CYP21A2P기인적동원성인물.선택4례선천성신상선피질증생증환자화5명정상대조,비교등위기인특이PCR인물여동원인물측서결과.결과 등위기인특이PCR인물확증화측서분석결과현시,4례환자균존재돌변,분별위IVS2 13A/C＞G、Arg356Trp화Arg149Pro,대조서렬정상.동원인물확증분석지능명학Arg149Pro돌변,환자화정상대조균존재IVS2-13A/C＞G화Arg356Trp돌변.결론 등위기인특이PCR기술결합측서가간편가고지진행CYP21A2기인돌변검측,구유림상응용추엄개치.
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.