无精子症因子微缺失拓展检测方法在2例性染色体拷贝数异常患者中的应用价值  

Application value of azoospermia factor microdeletion extended detection method in two special cases with abnormal sex chromosome copy numbers

作　　者：冯战启[1] 郭梁洁 苏俊祥 景治安[1] 李永乐 刘红彦 王红丹 Feng Zhanqi;Guo Liangjie;Su Junxiang;Jing Zhian;Li Yongle;Liu Hongyan;Wang Hongdan(Department of Urology,the First People's Hospital of Zhengzhou,Zhengzhou 450004,China;Medical Genetic Institute of Henan Province,People's Hospital of Zhengzhou University,Zhengzhou 450003,China;National Health Commission Key Laboratory of Birth Defects Prevention,Henan Key Laboratory of Population Defects Prevention,Henan Institute of Reproduction Health Science and Technology,Zhengzhou 450000,China)

机构地区：[1]郑州市第一人民医院泌尿外科,郑州450004 [2]郑州大学人民医院河南省人民医院医学遗传研究所,郑州450003 [3]国家卫生健康委出生缺陷预防重点实验室河南省人口缺陷干预技术研究重点实验室,河南省生殖健康科学技术研究院,郑州450000

出　　处：《中华生殖与避孕杂志》2022年第2期177-182,共6页Chinese Journal of Reproduction and Contraception

基　　金：国家自然科学基金(81501336);国家卫生健康委员会出生缺陷预防重点实验室开放课题(ZD202006);河南省科技攻关项目(202102310046);医学科技攻关项目(SBGJ202003001,LHGJ20200695)。

摘　　要：目的探讨Y染色体无精子症因子(azoospermia factor,AZF)微缺失拓展检测方法在遗传性不育和性发育异常疾病中的应用价值。方法本研究分别运用多重聚合酶链反应(polymerase chain reaction,PCR)结合琼脂糖凝胶电泳方法、复合荧光多重PCR毛细管电泳DNA片段分析技术以及染色体核型分析技术对2020年3月就诊于河南省人民医院生殖中心的1例不育症患者(患者1)和2020年4月就诊于河南省人民医院内分泌科的1例性腺发育异常儿童(患者2)进行检测。结果针对15个序列标签位点(sequence tagged site,STS)对这2例患者进行检测,结果未提示Y染色体AZF微缺失;针对27个STS遗传标记进行拓展检测,结果检测提示患者1位于X染色体长臂的STS位点扩增峰值是X染色体短臂扩增峰值的近3倍(Xqp),X染色体长臂的STR质控位点扩增峰为2个峰,且比值约为2∶1(GATA31E08和DXS6809),TAF9b位点在X染色体长臂扩增峰值与常染色体扩增峰值的比值约为3∶2,该患者X染色体长臂可能存在拷贝数异常。患者2,C03Yp、TAF9b、C01Yq和C11Xp位点在X染色体或Y染色体的扩增峰值与常染色体扩增峰值比值约为1∶1,X染色体的STR质控位点扩增峰为2个峰,且比值约为1∶1(GATA31E08和DXS6795),该患者可能存在X和Y染色体拷贝数异常;染色体核型分析显示患者1染色体核型为47,XY,i(X)(q10);患者2染色体核型为48,XXYY,与AZF微缺失拓展检测结果相印证。结论与传统AZF检测方法相比,本拓展检测方法不仅可以满足AZF检测需要,还可以提示性染色体拷贝数异常,较染色体核型分析技术,具有操作简捷的特点,可减低临床检验成本及工作量。Objective To explore the clinical application value of extended detection method for Y chromosome azoospermia factor(AZF)microdeletion in hereditary infertility and sexual development disorders.Methods Multiplex polymerase chain reaction(PCR)combined with agarose gel electrophoresis method,combined fluorescence multiplex PCR capillary electrophoresis DNA fragment analysis technique and chromosome karyotype analysis technique were used to detect an infertility patient who visited the Reproductive Center of Henan Provincial People's Hospital in March 2020(patient 1)and a child with sexual dysplasia who visited the Endocrinology Department of Henan Provincial People's Hospital in June 2020(patient 2).Results We found no AZF microdeletions on the Y-chromosome of the two patients to detect 15 sequence tagged site(STS)sequences.To detect the 27 genetic markers,it was found that in patient 1 the amplification peak of the STS locus on the long arm of the X chromosome was nearly three times as much as the amplification peak of the short arm of the X chromosome(Xqp),the STR quality control loci on the long arm of the X chromosome had two peaks,and the ratio was about 2∶1(GATA31E08 and DXS6809),and the ratio of the amplification peak of the long arm of the X chromosome to that of the autosome at the TAF9b locus was about 3∶2.Patient 1 might have an abnormal copy number of long arm of X chromosome.In patient 2,the ratio of the amplification peak of C03Yp,TAF9b,C01Yq and C11Xp on the X chromosome or Y chromosome to the amplification peak of autosomes was about 1∶1,and the amplification peak of the STR quality control site on the X chromosome was two peaks,and the ratio was about 1∶1(GATA31E08 and DXS6795).Patient 2 might have abnormal X and Y chromosome copy numbers.The results of karyotype analysis showed that the karyotype of patient 1 was 47,XY,i(X)(q10);the karyotype of patient 2 was 48,XXYY,which was consistent with the results of AZF microdeletion extension test.Conclusion Compared with the traditional AZF detec

关 键 词：Y染色体微缺失 男性不育 毛细管电泳 性染色体异常 

分 类 号：R698.2[医药卫生—泌尿科学]