摘要
目的应用微滴式数字化(droplet digital)PCR方法检测McCune-Albright综合征(MAS)患儿GNAS已知热点突变,并探讨其在MAS诊断中的应用价值。方法研究对象为122名就诊于上海交通大学医学院附属瑞金医院儿科的MAS患儿,针对MAS致病基因GNAS的已知突变热点(R201H/C),分别采用ddPCR技术、实时荧光焦磷酸解激活聚合反应技术(PAP)及二代测序方法检测其是否存在基因突变,并结合临床进行关联分析。结果122例MAS患儿中共89例完善了GNAS基因检测,发现存在突变者共57例。ddPCR、PAP及二代测序阳性率分别为77.42%、29.03%及56.25%。完善检测的经典三联征患者均检测出GNAS基因突变。其中,ddPCR方法在典型及不典型患儿外周血中阳性率分别为100%及73.1%,明显高于其余两种方法。性早熟合并骨病变患者GNAS突变检出率高于性早熟合并皮肤病变者,提示骨纤维结构不良伴性早熟是儿童MAS的临床诊断的重要依据。结论儿童MAS的内分泌表现以性早熟最为多见。骨纤维结构不良伴性早熟是临床诊断的重要权重因素。ddPCR灵敏度高,可为MAS的分子诊断提供帮助。
Objective To detect the known hotspot mutations of GNAS in children with McCune-Albrigtht syndrome(MAS)by droplet digital PCR,and to explore its application value in the diagnosis of MAS.Methods A total of 122 children with MAS were enrolled in the pediatric department of Ruijin Hospital Affiliated to Medical College of Shanghai Jiaotong University.For the known mutation hotspot of GNAS gene(R201H/C),dd-PCR,real-time fluorescent pyrophosphatic activation polymerase reaction(PAP)and second-generation sequencing were used to detect the presence of gene mutation and to analyse the relevance with the clinical features.Results GNAS gene mutation was detected in 89 out of 122 children with MAS and 57 cases were found to have mutations.The positive rates of ddPCR,PAP,and second generation sequencing were 77.42%,29.03%,and 56.25%,respectively.The GNAS gene mutation was detected in all classical triad patients.Among them,the positive rates of ddPCR in peripheral blood of typical and atypical children were 100%and 73.1%respectively,which were significantly higher than those of the other two methods.The detection rate of GNAS mutation in precocious puberty with bone lesions was higher than that in precocious puberty with skin lesions,suggesting that fibrous dysplasia with precocious puberty is an important basis for clinical diagnosis of MAS in children.Conclusion Precocious puberty is the most common endocrine manifestation of MAS in children.Bone fibrous dysplasia with precocious puberty is an important factor in clinical diagnosis.ddPCR has high sensitivity,which can be helpful for molecular diagnosis of MAS.
作者
谢轶雯
陆文丽
马晓宇
王伟
肖园
董治亚
王志敏
Xie Yiwen;Lu Wenli;Ma Xiaoyu;Wang Wei;Xiao Yuan;Dong Zhiya;Wang Zhimin(Department of Pediatrics,Ruijin Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200025,China)
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
2020年第5期416-420,共5页
Chinese Journal of Endocrinology and Metabolism
