为评估定量荧光PCR (QF-PCR)方法在男性不育遗传学诊断中的应用价值, 文章对78例非梗阻性男性不育患者, 采用精液常规检测精子情况, 并检测患者性激素水平; 采用QF-PCR方法对患者性染色体多态性STR位点及特异性位点进行检测; 采用常规染色体G显带方法进行核型分析; PCR检测AZF微缺失。结果显示78例非梗阻性男性不育患者中发现无精子症患者18例, 少精子症患者20例, 总检出率为48.72%。采用QF-PCR方法检出3例47, XXY患者, 2例46,XX(SRY+)性反转患者, 1例AZFc区微缺失患者, 与细胞培养染色体分析和AZF微缺失PCR检测结果相符。与传统方法相比, QF-PCR技术能更迅速、直接、可靠地检测到男性不育患者的染色体异常区域, 及早发现染色体细微结构异常, 有助于染色体异常造成的男性不育症的鉴别诊断。
To assess the clinical practice of quantitative fluorescence PCR (QF-PCR) in genetic diagnosis of male infertility patients, 78 nonobstructive male infertility patients were pooled for semen routine screening and sexual hormone determination; QF-PCR was applied to detect the polymorphic short tandem repeat (STR) and specific sequence tagged site (STS) of sex chromosomes; routine chromosome G-band was used for karyotype analysis and PCR was used for the detection of AZF microdeletion. Routine screening of semen found 18 azoospermia and 20 oligospermia patients (48.72%). Three patients with 47, XXY, two with 46,XX(SRY+)and one with AZFc microdeletion were detected using QF-PCR technique which were verified by chromosome G-band and PCR. This study suggests that QF-PCR is a compre-hensive, rapid and reliable method for detecting abnormal chromosomal regions and microstructures compared with tradi-tional tests and provides a better candidate for diagnosis of male infertility caused by chromosomal anomalies and gene mutation.
[1] Rowe PJ, Comhaire FH, Hargreave TB, Mahmoud AMA. 世界卫生组织男性不育标准化检查与诊疗手册. 李铮, 张忠平, 黄翼然, 王益鑫, 朱晓斌, 刘毅东, 张斌, 卢慧, 王磊, 曹小蓉, 刘勇, 王一飞, 译. 北京: 人民卫生出版社, 2007: 1-2.
[2] 世界卫生组织. 人类精液及精子-宫颈粘液相互作用实验室检验手册 (第4版). 谷翊群, 陈振文, 于和鸣, 罗宏志, 裴开颜, 译. 北京: 人民卫生出版社, 2001: 45.
[3] Dada R, Gupta NP, Kucheria K. Molecular screening for Yq microdeletion in men with idiopathic oligozoospermia and azoospermia. J Biosci, 2003, 28(2): 163-168.
[4] Cirigliano V, Voglino G, Ordoñez E, Marongiu A, Paz Cañadas M, Ejarque M, Rueda L, Lloveras E, Fuster C, Adinolfi M. Rapid prenatal diagnosis of common chromosome aneuploidies by QF-PCR, results of 9 years of clinical experience. Prenat Diagn, 2009, 29(1): 40-49.
[5] Liao C, Yang X, Li FT, Li J, Li DZ. The detection of aneuploidy and maternal contamination by QF-PCR in sam-ples undergoing prenatal diagnosis for thalassemia in Southern China. Eur J Obstet Gynecol Reprod Biol, 2009, 144(2): 149-152.
[6] Onay H, Ugurlu T, Aykut A, Pehlivan S, Inal M, Tinar S, Ozkinay C, Ozkinay F. Rapid prenatal diagnosis of common aneuploidies in amniotic fluid using quantitative fluorescent polymerase chain reaction. Gynecol Obstet Invest, 2008, 66(2): 104-110.
[7] Badenas C, Rodríguez-Revenga L, Morales C, Mediano C, Plaja A, Pérez-Iribarne MM, Soler A, Clusellas N, Borrell A, Sánchez MÁ, Miró E, Sánchez A, Milà M, Jiménez W. Assessment of QF-PCR as the first approach in prenatal diagnosis. J Mol Diagn, 2010, 12(6): 828-834.
[8] Christopoulou S, Christopoulou G, Hatzaki A, Hatzi-pouliou A, Donoghue J, Karkaletsi M, Kaminopetros P, Sifakis S, Velissariou V. The replacement of cytogenetic analysis by direct chorionic villi sampling preparation with quantitative fluorescence PCR. Gynecol Obstet Invest, 2009, 68(4): 255-261.
[9] Morales C, Sánchez A, Bruguera J, Margarit E, Borrell A, Borobio V, Soler A. Cytogenetic study of spontaneous abortions using semi-direct analysis of chorionic villi samples detects the broadest spectrum of chromosome abnormalities. Am J Med Genet A, 2008, 146A(1): 66-70.
[10] 卢彦平, 程静, 姜淑芳, 张利文, 高志英, 韩冰, 袁慧军, 李亚里. 多重荧光定量PCR技术快速诊断21三体及18三体方法的建立及临床应用. 遗传, 2010, 32(11): 1141-1146.
[11] Tiepolo L, Zuffardi O. Localization of factors controlling spermatogenesis in the nonfluorescent portion of the hu-man Y chromosome long arm. Hum Genet, 1976, 34(2): 119-124.
