Screening of azoospermia factor microdeletions on Y chromosome in infertile men by QF-PCR
Received date: 2013-12-17
Revised date: 2014-02-12
Online published: 2014-05-28
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.
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 . DOI: 10.3724/SP.J.1005.2014.0552
[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
/
| 〈 |
|
〉 |
www.chinagene.cn
备案号：京ICP备09063187号
