1例染色体结构变异无精症患者的纳米孔测序临床应用  

Clinical application of nanopore sequencing in an azoospermia patient with chromosome structural variation

作　　者：王亚 黄明涛 刘安 周冉[1] 张沁欣 胡平[1] 许争峰[1] WANG Ya;HUANG Mingtao;LIU An;ZHOU Ran;ZHANG Qinxin;HU Ping;XU Zhengfeng(Department of Prenatal Diagnosis,State Key Laboratory of Reproductive Medicine,Women′s Hospital of Nanjing Medical University,Nanjing Maternity and Child Health Care Hospital,Nanjing 210001,Jiangsu,China)

机构地区：[1]南京医科大学附属妇产医院生殖医学国家重点试验室,南京市妇幼保健院产前诊断中心,南京210001

出　　处：《临床检验杂志》2022年第3期220-224,共5页Chinese Journal of Clinical Laboratory Science

基　　金：国家重点研发开发项目(2018YFC1002402);国家自然科学基金(81971398)。

摘　　要：目的评价牛津纳米孔测序技术(oxford nanopore technologies,ONT)在诊断复杂结构变异中的临床应用价值。方法采集1例复杂结构变异的非梗阻性无精症患者外周血样本,以染色体G显带核型分析(karyotype analysis)与染色体微阵列分析(Chromosomal microarray analysis,CMA)为参考,进行纳米孔测序寻找致病原因,评价纳米孔测序对变异的检出能力。结果在该患者的基因组中发现,CMA检测出4个缺失,5个重复,共计9个临床意义不明的染色体拷贝数变异(VOUS),3例纯合子状态区段(regions of homozygosity,ROH)以及15q15.3区段缺失,内含PPIP5K1、CKMT1B、STRC、CATSPER2基因;纳米孔测序共检测出6260个缺失、5624个插入、115个重复、22个倒位、32个易位,共计12053个结构变异,能够注释5559个结构变异影响2406个基因;纳米孔测序鉴定出平衡易位46,XY,t(1;3)(q22;q28)与染色体核型结果46,XY,t(1;3)(q21;q27)基本一致,在Chr1:156359988-156363186区域存在3198 bp缺失,该区域影响的基因有TSACC、RHBG和FGF12。结论纳米孔测序技术可快速、高效、准确检测人外周血标本中染色体复杂结构变异和受影响的基因,可检测出传统检测方法无法检出的缺失片段,是常规方法的补充,具有重要的临床应用价值。Objective To evaluate the application value of oxford nanopore technology(ONT)in the diagnosis of complex structural variations(SVs).Methods The peripheral blood sample of a patient with non-obstructive azoospermia and complex structural variations was collected,and identified by the chromosome G-banding karyotype analysis and chromosomal microarray analysis(CMA),which were taken as the reference methods.Meanwhile,the sample was detected by the ONT to discover the cause of disease,and the detection ability of ONT for variations was evaluated.Results A total of 9 copy number variations of uncertain significance(VOUS),including 4 deletions and 5 duplications,3 regions of homozygosity(ROH)and 15q15.3(containing PPIP5K1,CKMT1B,STRC and CATSPER2 genes)deletion were identified by the CMA.A total of 12053 structural variations,including 6260 deletions,5624 insertions,115 duplications,22 inversions and 32 translocations,were identified by the ONT,which could annotate 5559 structural variations and affect 2406 genes.The balanced translocation of 46,XY,t(1;3)(q22;q28)detected by the ONT was basically consistent with the result of karyotype analysis[46,XY,t(1;3)(q22;q28)].In addition,a 3198 bp deletion in the Chr1:156359988-156363186 region was detected,which could affect TSACC,RHBG and FGF12 genes.Conclusion The ONT can rapidly,efficiently and accurately detect the complex structural variations of chromosomes and affected genes in human peripheral blood samples,and deletions that cannot be detected by traditional detection methods,which is a supplement to traditional methods and has important clinical application value.

关 键 词：纳米孔测序 结构变异 基因 

分 类 号：R446[医药卫生—诊断学]