杜氏肌营养不良新生儿筛查及诊断策略研究  

Study on newborn screening for Duchenne muscular dystrophy and diagnostic strategy

作　　者：鲍幼维 潘小莉[1] 潘婕文[1] 潘澍青 庄丹燕[1] 李海波 曹宗富 Bao Youwei;Pan Xiaoli;Pan Jiewen;Pan Shuqing;Zhuang Danyan;Li Haibo;Cao Zongfu(Ningbo Women and Children’s Hospital,Ningbo Municipal Key Laboratory of Comprehensive Prevention and Treatment of Birth Defects,Ningbo,Zhejiang 315012,China;National Research Institute for Family Planning,Beijing 100081,China)

机构地区：[1]宁波市妇女儿童医院出生缺陷综合防治实验室,315012 [2]国家卫生计生委科学技术研究所,北京100081

出　　处：《中华医学遗传学杂志》2021年第5期430-434,共5页Chinese Journal of Medical Genetics

基　　金：浙江省医药卫生科技计划项目(2020KY889);宁波市科技创新团队(2014B82003);国家人口与生殖健康科学数据中心(2005DKA32408);宁波市社会发展公益项目(2019C50070)。

摘　　要：目的探究将肌型肌酸激酶同工酶(MM isoenzyme of creatine kinase,CK-MM)检测用于筛查新生儿杜氏肌营养不良症(Duchenne muscular dystrophy,DMD)的可行性。方法对10252例男性新生儿干血斑的CK-MM含量进行测定,将结果按孕周、采血时间、实验间隔时间分组,并分析需要进行基因诊断的CK-MM截断值,对超过截断值者用多重连接探针扩增及二代测序进行确诊。结果非参数秩和检验分析提示新生儿CK-MM值浓度中位数随孕周增加而上升,随采血年龄增加而下降,且干血斑中的CK-MM在2~8℃的干燥环境中可稳定14天。通过对10252名新生儿的CK-MM值进行分析,设定截断值为700 ng/mL,并在超过截断值的4例新生儿中确诊2例阳性病例。二者均携带外显子缺失,其CK-MM值均大于2000 ng/mL。结论检测干血斑中CK-MM的含量是筛查新生儿DMD基因突变的有效手段,其操作简单,成本低廉,可用于大规模筛查,对于DMD患儿的早期干预与治疗具有重要的意义。Objective To establish a newborn screening system for Duchenne muscular dystrophy(DMD)through assessment of MM isoenzyme of creatine kinase(CK-MM)activity.Methods The CK-MM level was detected using dry blood spot filter paper from 10252 male newborns.The results were grouped based on their gestational age,sampling time and intervals between the experiments.The threshold value for CK-MM necessitating genetic testing was determined.Next-generation sequencing(NGS)was carried out for those with a CK-MM value over the threshold,and the result was verified by multiplex ligation-dependent probe amplification(MLPA).Results Based on the result of non-parametric rank sum test,the median CK-MM concentration has increased with the gestational age,and was inversely correlated with the age of the newborns among unaffected specimens.CK-MM on dry blood spot filter paper can be stable for 14 days at 2-8℃.Statistical analysis of CK-MM value of the 10252 neonates suggested that the threshold may be set as 700 ng/mL.Exonic deletions were found in 2 confirmed cases,whose CK-MM level was greater than 2000 ng/mL.Conclusion Detection of CK-MM in dry blood spot filter paper has provided an effective method for newborn screening of DMD.This simple and inexpensive method can be used for large-scale screening,which is of great value to the early intervention and treatment of the disease.

关 键 词：杜氏肌营养不良症 新生儿筛查 基因突变 肌酸激酶同工酶 

分 类 号：R722.1[医药卫生—儿科]