酶切法检测6-丙酮酰基四氢蝶呤合成酶基因常见突变  

Detection of the prevalent mutations of 6-pyruvoyl-tetrahydropterin synthase gene by PCR-RFLP analysis in Chinese patients

作　　者：瞿宇晋[1] 宋昉[1] 王红[1] 金煜炜[1] 

机构地区：[1]首都儿科研究所遗传室,北京100020

出　　处：《中华医学遗传学杂志》2006年第6期680-682,共3页Chinese Journal of Medical Genetics

基　　金：国家十五科技攻关项目基金(2004BA720A04)~~

摘　　要：目的建立一种快速、特异的限制性内切酶酶切分析法检测中国人6-丙酮酰基四氢蝶呤合成酶(6-pyruvoyl-tetrahydropterinsynthase,PTPS)基因的常见突变。方法应用人工构建的酶切位点,对4例四氢生物喋呤缺乏症(tetrahydrobiopterindeficiency,BH4D)患儿及其父母进行限制性片段长度多态性(restric-tionfragmentlengthpolymorphism,RFLP)分析,检测中国人最常见的3种PTPS基因突变N52S、P87S和D96N;同时采用测序的方法验证PCR-RFLP的结果。结果4例BH4D患儿的PTPS突变基因型为N52S/-、P87S/D96N、N52S/D96N和D96N/-;PCR-RFLP分析与DNA测序结果一致。结论(1)人工引入酶切位点的酶切方法操作简便、特异性好,适于PTPS基因常见突变的临床检测;⑵高苯丙氨酸血症(hyperpheny-lalaninemia,HPA)患儿有必要进一步分析PTPS基因,以便早期鉴别BH4D,选择正确的治疗方案。Objective To establish a single and rapid method for detecting the prevalent mutations of 6-pyruvoyl-tetrahydropterin synthase （PTPS） gene in Chinese patients with 6-pyruvoyl-tetrahydropterin synthase deficiency （FTPSD）. Methods PCR-restriction fragment length polymorphism （PCR-RFLP） was used to detect three prevalent PTPS gene mutations in 4 cases of tetrahydrobiopterin defiency （BH4D） and their parents, which was performed by the artificial construct restriction site （ACRS）. PCR-RFLP included Hpa Ⅰ digestion for the detection of mutation N52S （155A 〉 G）, Hae Ⅲ for P87S（259C 〉 T） and EcoR Ⅰ for D96N（286G 〉 A）. The mutations of PTPS gene were confinned by direct sequencing. Results The genotypes of 4 PTPS deficient patients were identified： N52S/-, P87S/D96N, N52S/D96N and D96N/-. The result of direct sequencing was coincident with that of PCR-RFLP analysis. Conclusion （1） The PCR-RFLP analysis formed by ACRS would be a good way for detecting the three prevalent PTPS gene mutations in Chinese patients with PTPSD. （2） The PTPS gene analysis is very important for all patients with hyperphenylalaninemia, which would be useful for early clinical diagnosis and correct treatment of BH4D patients.

关 键 词：四氢生物蝶呤缺乏症 6-丙酮酰基四氢蝶呤合成酶基因 突变检测 聚合酶链反应-限制性片段长度多态性 

分 类 号：R450[医药卫生—治疗学]