CNVPLUS■微阵列芯片在DMD基因诊断中的应用  

Applications of CNVPLUS■-array in diagnosis of DMD gene

作　　者：郭彩琴[1,2] 方丹枫 杨婷婷 刘怡 朱佳依 余永国 Guo Caiqin;Fang Danfeng;Yang Tingting;Liu Yi;Zhu Jiayi;Yu Yongguo(Department of Medical Genetics and Prenatal Diagnosis,Wuxi Maternity and Child Health Care Hospital(Affiliated Women′s Hospital of Jiangnan University),Wuxi 214002,China;Department of Pediatric Endocrinology and Genetics,Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China;Department of Pediatric Internal Medcine,Taizhou Central Hospital(Taizhou University Hospital),Taizhou 318000,China)

机构地区：[1]无锡市妇幼保健院(江南大学附属妇产医院)医学遗传与产前诊断科,无锡214002 [2]上海交通大学医学院附属新华医院儿内分泌遗传科,上海200092 [3]台州市中心医院(台州学院附属医院)儿内科,台州318000

出　　处：《中华预防医学杂志》2024年第4期508-515,共8页Chinese Journal of Preventive Medicine

基　　金：国家重点研发计划(2022YFC2703400)。

摘　　要：探讨CNVPLUS■微阵列芯片(CNVPLUS■-array)在 DMD基因诊断中的应用价值。选取2014年1月至2023年3月在上海交通大学医学院附属新华医院儿内分泌遗传科就诊、临床诊断Duchenne/Becker肌营养不良(DMD/BMD)的96例儿童纳入回顾性分析,分别采用CNVPLUS■-array技术和多重连接探针扩增(MLPA)-二代测序(NGS)-Sanger测序序贯法检测患儿外周血 DMD基因。序贯法中,MLPA检出的单外显子缺失运用聚合酶链式反应(PCR)验证,PCR结果正常的样本再行Sanger测序;MLPA阴性的样本行NGS检测并用Sanger测序验证。结果显示,96例样本中,CNVPLUS■-array检出 DMD基因致病性变异91例,其中大片段缺失/重复即拷贝数变异(CNV)76例、微小变异即单核苷酸变异/小插入缺失(SNV/Indel)15例,5 d完成所有样本的实验和诊断;序贯法则检出致病性CNV 76例、SNV/Indel 20例,48 d完成全部实验和诊断。5例CNVPLUS■-array漏诊的SNV/Indel样本优化运算模式后致病位点可以正确聚类。综上,CNVPLUS■-array由于集CNV和SNV探针为一体,能同步检测 DMD基因的CNV和SNV/Indel,但对CNV的诊断性能优于SNV/Indel,有待优化运算模式必要时需补充其他检测以减少漏诊风险。To explore the value of CNVPLUS■-array in the diagnosis of the DMD gene.A retrospective study was performed on 96 children who were clinically diagnosed with Duchenne or Becker muscular dystrophies(DMD/BMD)at the Department of Pediatric Endocrinology and Genetics of Xinhua Hospital affiliated to Shanghai Jiaotong University School of Medicine from January 2014 to March 2023.DNA was extracted from these children′s peripheral blood and divided into two parts.Variations of the DMD gene were detected by using CNVPLUS■-array and sequential testing of MLPA—NGS—Sanger.In the sequential method,single exon deletions detected by MLPA were first verified by polymerase chain reaction(PCR)and then were tested by Sanger′s sequencing if PCR results were normal.The results showed that,among 96 samples,91 cases with the pathogenic variation of the DMD gene were detected by the CNVPLUS■-array,including 76 cases with large deletion/duplication(copy number variation,CNV)and 15 cases with small variation(single nucleotide variant or small insertion/deletion,SNV/Indel).All samples were tested and diagnosed within 5 days.In contrast,76 cases with pathogenic CNV and 20 cases with pathogenic SNV/Indel were detected in the DMD gene by sequential method.However,all of the experiments and diagnoses were completed within 48 days.Moreover,5 cases with SNV/Indel in the DMD gene were correctly clustered after the operation mode was optimized.In summary,as a new micro-array integrating CNV and SNV probes,CNVPLUS■-array can detect CNV and SNV/Indel in the DMD gene simultaneously while the application of CNVPLUS■-array could save a lot of time and manpower.CNVPLUS■-array had an excellent diagnostic performance for CNV of the DMD gene.As for SNV/Indel,the diagnostic performance was slightly poor and the operation mode should be optimized.If necessary,other testing technologies should be supplemented to reduce the risk of missed diagnosis.

关 键 词：DUCHENNE肌营养不良 Becker肌营养不良 DMD基因 CNVPLUS■微阵列芯片 

分 类 号：R746.2[医药卫生—神经病学与精神病学] R440[医药卫生—临床医学]