QF-PCR筛查男性不育患者Y染色体无精子症因子微缺失

doi:10.3724/SP.J.1005.2014.0552

遗传 ›› 2014, Vol. 36 ›› Issue (6): 552-557.doi: 10.3724/SP.J.1005.2014.0552

QF-PCR筛查男性不育患者Y染色体无精子症因子微缺失

张媛媛¹, 杜强², 刘晓亮¹, 崔婉婷¹, 何蓉¹, 赵彦艳¹

1. 中国医科大学附属盛京医院临床遗传科, 沈阳 110004;
2. 中国医科大学附属盛京医院生殖中心, 沈阳 110004

收稿日期:2013-12-17 修回日期:2014-02-12 出版日期:2014-06-20 发布日期:2014-05-28
通讯作者: 赵彦艳,博士,教授,研究方向：复杂疾病的分子遗传学。E-mail:yyzhao@sj-hospital.org E-mail:zyy.0526@163.com
作者简介:张媛媛,博士,研究方向：复杂疾病的分子遗传学。Tel:024-96615-75311;E-mail:zyy.0526@163.com
基金资助:
国家自然科学基金项目(编号：81270343,81100187)资助

Screening of azoospermia factor microdeletions on Y chromosome in infertile men by QF-PCR

Yuanyuan Zhang¹, Qiang Du², Xiaoliang Liu¹, Wanting Cui¹, Rong He¹, Yanyan Zhao¹

1. Department of Clinical Genetics, Shengjing Hospital, China Medical University, Shenyang 110004, China;
2. Center of Reproductive Medicine, Shengjing Hospital, China Medical University, Shenyang 110004, China

Received:2013-12-17 Revised:2014-02-12 Online:2014-06-20 Published:2014-05-28

摘要/Abstract

摘要：

为评估定量荧光PCR(Quantitative fluorescent polymerase chain reaction, QF-PCR)技术在快速筛查无精子症因子(Azoospermia factor, AZF)微缺失中的应用, 文章对1218例非梗阻性无精子症、少精子症的男性不育患者, 采用多重QF-PCR结合毛细管电泳技术, 检测Y染色体长臂AZF区9个序列标签位点(Sequence tagged site, STS)以及性染色体短臂的AMEL(Amelogenin)和SRY(Sex-determining region of Y chromosome)位点, 辅以常规染色体G显带方法进行核型分析。结果显示, 1218例患者中105例可见AZF区微缺失(8.62%), 其中AZFc区缺失(67.62%)最常见, 其次为AZFb,c区缺失(20.95%); AZFb区缺失(7.62%)和AZFa区缺失(3.81%)则较少见; 另有5例患者为AZFa,b,c区缺失合并AMEL-Y缺失, 提示可能缺少Y染色体, 经核型分析验证为46,XX(性反转)。105例AZF区微缺失患者的染色体核型分析显示染色体异常16例, 其中“Yqh-”12例。根据AMEL-X/AMEL-Y比值, 可见1218例患者中86例可能存在性染色体异常, 经核型分析验证, 68例为性染色体非整倍体。多重QF-PCR技术, 一个反应即能检测样本的多个位点, 并可提示性染色体是否存在异常, 有助于男性不育患者尽早明确病因, 也为后续的检查和治疗提供依据。

关键词: 无精子症因子, 微缺失, Y染色体, 定量荧光PCR, 男性不育症

Abstract:

To assess the application of quantitative fluorescent polymerase chain reaction (QF-PCR) on rapid screening of azoospermia factor (AZF) microdeletions, 1218 infertile men with non-obstructive azoospermia or oligospermia were detected for 9 sequence tagged sites (STSs) in AZF region by multiplex QF-PCR combined with capillary electrophoresis. AMEL (amelogenin) as well as SRY (sex-determining region of Y chromosome) located on short arm of sex chromosome was selected as internal control. Karyotyping was performed on Giemsa-banded metaphase chromosomes of peripheral blood lymphocytes. Of the 1218 patients, 105 (8.62%) were identified as AZF microdeletions. Deletion of AZFc (67.62%) was the most frequent, followed by deletion of AZFb,c (20.95%), AZFb (7.62%) and AZFa (3.81%). Five patients presented with deletions of both AZFa,b,c and AMEL-Y, indicating sex reversal which was confirmed to be 46,XX by karyotyping. Among the 105 patients with AZF microdeletions, 16 were karyotyped as chromosomal anomalies, most commonly 46,XY,Yqh- (75%, 12/16). In addition, of the total 1218 patients examined, 86 patients showed abnormal AMEL-X/AMEL- Y ratio, suggesting a possibility of sex chromosome anomalies, and 68 of them were verified as sex chromosome aneuploid by karyotyping. Multiplex QF-PCR is capable to detect all markers in one reaction and is also suggestive for sex chromosome anomalies. It could serve as an effective technique for screening Y-microdeletions, and thus have general application in diagnosis and treatment of male infertility.

Key words: azoospermia factor, microdeletion, Y chromosome, quantitative fluorescent polymerase chain reaction, male infertility

张媛媛, 杜强, 刘晓亮, 崔婉婷, 何蓉, 赵彦艳. QF-PCR筛查男性不育患者Y染色体无精子症因子微缺失[J]. 遗传, 2014, 36(6): 552-557.

Yuanyuan Zhang, Qiang Du, Xiaoliang Liu, Wanting Cui, Rong He, Yanyan Zhao. Screening of azoospermia factor microdeletions on Y chromosome in infertile men by QF-PCR[J]. HEREDITAS(Beijing), 2014, 36(6): 552-557.

参考文献

[1]Dohle GR, Colpi GM, Hargreave TB, Papp GK, Jungwirth A, Weidner W. EAU guidelines on male infertility。 Eur Urol, 2005, 48(5): 703-711.
[2]葛少钦, Grifin J, 刘丽华, Aston KI, Simon L, Jenkins TG , Emery BR , Carrell DT. 特发性少精子症和无精子症与Pygo2基因蛋白编码区SNPs 的相关性. 遗传, 2013, 35(5): 616-622.
[3]冉静, 韩婷婷, 丁显平, 魏霞, 张丽媛, 张玉平, 李天俊, 聂双双, 陈林. Y 染色体单倍群与西南地区男性生精障碍的相关性. 遗传, 2013, 35(1): 73-78.
[4]阮健, 杜卫东. 男性不育与基因缺陷. 遗传, 2010, 32(5): 411-422.
[5]Kim MJ, Choi HW, Park SY, Song IO, Seo JT, Lee HS. Molecular and cytogenetic studies of 101 infertile men with microdeletions of Y chromosome in 1, 306 infertile Korean men. J Assist Reprod Genet, 2012, 29(6): 539- 546.
[6]齐漫龙, 张媛媛, 刘晓亮, 何蓉, 赵彦艳. QF-PCR 在染色体异常男性不育症诊断中的应用. 遗传, 2011, 33(8): 895-900.
[7]World Health Organization. WHO laboratory manual for the examination and processing of human semen. 5th ed. World Health Organization Press, 2010.
[8]Krausz C, Hoefsloot L, Simoni M , Tuttelmann F. EAA/ EMQN best practice guidelines for molecular diagnosis of y-chromosomal microdeletions. State of the art 2004. Int J Androl, 2004, 27(4): 240-249.
[9]Tiepolo L, Zuffardi O. Localization of factors controlling spermatogenesis in the nonfluorescent portion of the human Y chromosome long arm. Hum Genet, 1976, 34(2): 119-124.
[10]Vogt PH, Edelmann A, Kirsch S, Henegariu O, Hirschmann P, Kiesewetter F, Köhn FM, Schill WB, Farah S, Ramos C, Hartmann M, Hartschuh W, Meschede D, Behre HM, Castel A, Nieschlag E, Weidner W, Gröne HJ, Jung A, Engel W, Haidl G. Human Y chromosome azoo-spermia factors (AZF) mapped to different subregions in Yq11. Hum Mol Genet, 1996, 5(7): 933-943.
[11]Hofherr SE, Wiktor AE, Kipp BR, Dawson DB, Van Dyke DL. Clinical diagnostic testing for the cytogenetic and molecular causes of male infertility: the Mayo Clinic experience. J Assist Reprod Genet, 2011, 28(11): 1091-1098.
[12]Zhang YS, Dai RL, Wang RX, Zhang HG, Chen S, Liu RZ. Analysis of Y chromosome microdeletion in 1738 infertile men from northeastern China. Urology, 2013, 82(3): 584- 588.
[13]Wu Q, Chen GW, Yan TF, Wang H, Liu YL, Li Z, Duan SW, Sun F, Feng Y, Shi HJ. Prevalent false positives of azoospermia factor a (AZFa) microdeletions caused by single-nucleotide polymorphism rs72609647 in the sY84 screening of male infertility. Asian J Androl, 2011, 13(6): 877-880.
[14]Wang RX, Fu C, Yang YP, Han RR, Dong Y, Dai RL, Liu RZ. Male infertility in China: laboratory finding for AZF microdeletions and chromosomal abnormalities in infertile men from Northeastern China. J Assist Reprod Genet, 2010, 27(7): 391-396.
[15]付莉, 丁显平, 沈梦婕, 李创, 聂双双, 权强. 1011例男性不育患者Y染色体无精子症因子微缺失的筛查与临床表型分析. 中华医学遗传学杂志, 2012, 29(2): 184- 187.
[16]Hopps CV, Mielnik A, Goldstein M, Palermo GD, Rosenwaks Z, Schlegel PN. Detection of sperm in men with Y chromosome microdeletions of the AZFa, AZFb and AZFc regions. Hum Reprod, 2003, 18(8): 1660-1665.
[17]Reijo R, Lee TY, Salo P, Alagappan R, Brown LG, Rosenberg M, Rozen S, Jaffe T, Straus D, Hovatta O, de la Chapelle A, Silber S, Page DC. Diverse spermatogenic defects in humans caused by Y chromosome deletions encompassing a novel RNA-binding protein gene. Nat Genet, 1995, 10(4): 383-393.
[18]Ferlin A, Arredi B, Speltra E, Cazzadore C, Selice R, Garolla A, Lenzi A, Foresta C. Molecular and clinical characterization of Y chromosome microdeletions in infertile men: a 10-year experience in Italy. J Clin Endocrinol Metab, 2007, 92(3): 762-770.
[19]Demir B, Arikan II, Bozdag G, Esinler I, Karakoc Sokmensuer L, Gunalp S. ICSI outcome of patients with severe oligospermia vs. non-obstructive azoospermia. Clin Exp Obstet Gynecol, 2012, 39(2): 141-143.
[20]Murphy KM, Co

编辑推荐

Metrics

www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:

QF-PCR筛查男性不育患者Y染色体无精子症因子微缺失

Screening of azoospermia factor microdeletions on Y chromosome in infertile men by QF-PCR

PDF (PC)

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	王飞, 王萌, 张兴华, 宇克莉, 郑连斌, 杨亚军. 中国西南地区3个隔离人群遗传亚结构分析[J]. 遗传, 2022, 44(5): 424-431.
[2]	王燕超,马晓燕,孙筱放,冼嘉嘉,李少英,何文智,王晓蔓,黎青. Y染色体微缺失人群中Y-STR等位基因缺失模式分析[J]. 遗传, 2019, 41(3): 243-253.
[3]	田娇阳, 李玉春, 孔庆鹏, 张亚平. 遗传学视角下东亚人群的起源和演化[J]. 遗传, 2018, 40(10): 814-824.
[4]	杨仙荣, 王美琴, 李少华. 人类Y染色体的演化[J]. 遗传, 2014, 36(9): 849-856.
[5]	王跃力, 叶峻杰, 李宗芳, 郑水, 马丽, 郭海, 杨丽娟, 程宝文. 对法医学相关的DYS549、DYS527和DYS459基因座在男性不育症人群中的缺失、重复调查[J]. 遗传, 2014, 36(8): 786-792.
[6]	文少卿谢小冬徐丹. 接触与混合——从Y染色体的角度看东乡人群及其语言的关系[J]. 遗传, 2013, 35(6): 761-770.
[7]	马志杰，钟金城，韩建林，徐惊涛，刘仲娜，白文林. 牦牛分子遗传多样性研究进展[J]. 遗传, 2013, 35(2): 151-160.
[8]	冉静，韩婷婷，丁显平，魏霞，张丽媛，张玉平，李天俊，聂双双，陈林. Y染色体单倍群与西南地区男性生精障碍的相关性[J]. 遗传, 2013, 35(1): 73-78.
[9]	百茹峰，杨利海，袁丽，梁权赠，鲁涤，杨雪，石美森. 福建畲族群体17个Y-STR基因座单倍型及遗传关系[J]. 遗传, 2012, 34(8): 1020-1030.
[10]	石美森，百茹峰，傅博. 山西汉族17个Y-STR基因座遗传多态性及遗传关系[J]. 遗传, 2011, 33(3): 228-238.
[11]	张晓明，乐祥鹏，张春梅，蓝贤勇，陈宏，雷初朝. 沼泽型水牛Y染色体微卫星标记的筛选与多态性检测[J]. 遗传, 2010, 32(3): 242-247.
[12]	冯冬亮，刘长晖，梁祚仁，刘超 . 广西4个少数民族17个Y-STR基因座的多态性分析[J]. 遗传, 2009, 31(9): 921-935.
[13]	张永科，陈争，范安，张亚男，武艳平，赵倩君，周璨林，毛新民，藏玉亮，叶尔哈孜·扎尔胡马尔，哈森别克·马力克，哈布德力·达拉拜依，卡马力亚·托塔哈孜，梁玲，古力努尔·叶尔肯，马月辉，饶绍奇. 新疆阿勒泰地区图瓦人与邻近人群遗传关系初探[J]. 遗传, 2009, 31(8): 818-824.
[14]	石美森，百茹峰，于晓军，唐剑频 . 广东汉族22个Y-STR基因座遗传多态性及遗传关系分析[J]. 遗传, 2008, 30(9): 1136-1142.
[15]	石美森，百茹峰，张金生，于晓军. 辽宁满族11个Y-STR基因座多态性及遗传关系的分析[J]. 遗传, 2008, 30(5): 583-589.