目标序列捕获二代测序技术在苯丙酮尿症基因诊断中的应用  

Application of captured high throughput sequencing in the genetic diagnosis of phenylketonuria

作　　者：陈瑛[1] 魏晓明 王本敬[3] 易鑫 毛君[4] 

机构地区：[1]南京医科大学附属苏州医院新生儿科-中心实验室,苏州215002 [2]深圳华大基因研究院,深圳518083 [3]南京医科大学附属苏州医院妇科,苏州215002 [4]南京医科大学附属苏州医院产科,苏州215002

出　　处：《复旦学报（医学版）》2015年第3期403-408,共6页Fudan University Journal of Medical Sciences

基　　金：江苏省"十二五"科教兴卫工程医学重点人才资助项目(RC2011036);苏州市生殖与遗传转化医学重点实验室建设项目(SZS201206)~~

摘　　要：目的评估目标序列捕获二代测序技术在苯丙酮尿症(phenylketonuria,PKU)基因诊断中的应用价值。方法采用目标区序列捕获及第二代测序技术对9例经典型PKU患者的苯丙氨酸羟化酶(phenylalanine hydroxylase,PAH)基因进行检测;采用Sanger测序技术对患者及其父母基因突变型进行验证。结果 9例患者检测到PAH基因的12种致病性突变,包括7种错义突变(p.I65T、p.F161S、p.Q204C、p.R241C、p.L242F、p.R243Q和p.Q375E),2种无义突变(p.R111X和p.Y356X),3种剪接突变[c.442-1G>A(IVS4-1G>A)、c.1315+6T>A(IVS12+6T>A)和c.1316-2A>C(IVS12-2A>C)],以及3种非致病性的变异(p.Q232Q、p.V245V和p.L385L),致病性突变均来自患者父母。其中,2个致病性突变未见报道,分别为c.1316-2A>C和p.Q375E(CAA->GAA)。结论目标序列捕获二代测序可以准确检测出PAH基因突变,对遗传病因明确的疾病具有一定的临床应用价值。Objective Assess the performance of captured high throughput sequencing in the genetic diagnosis of phenylketonuria （PKU）.Methods Target region sequence capture and next-generation sequencing were used to detect phenylalanine hydroxylase （PAH） gene of 9 children affected with typical PKU.Sanger sequencing was used to confirm the results of the patients and their parents.Results Nine children carried 12 pathogenicity mutations in PAH gene,including 7 missense mutation （p.I65T,p.F161S,p.Q204C,p.R241C,p.L242F,p.R243Q and p.Q375E）,2 nonsense mutation （p.R111X and p.Y356X）,3 splicing mutation[c.442-1G＞A（IVS4-1G＞A） 、c.1315 ＋ 6T＞A（IVS12 ＋ 6T ＞A） and c.1316-2A＞C（IVS12-2A＞C）] and 3 unpathologic silence mutations （p.Q232Q,p.V245V and p.L385L）.All the mutations were from their parents,in which 2 novel pathogenicity mutations were not reported before,including c.1316-2A＞C and p.Q375E （CAA-＞GAA）.Conclusions Next generation sequencing technology is a useful tool for the detection of PAH gene,which is valuable for diagnosing diseases of genetic etiology confirmed in clinical application.

关 键 词：苯丙酮尿症 苯丙氨酸羟化酶 序列捕获 第二代测序技术 

分 类 号：R596.2[医药卫生—内科学] R394[医药卫生—临床医学]