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

作　　者：张媛媛[1] 杜强[2] 刘晓亮[1] 崔婉婷[1] 何蓉[1] 赵彦艳[1] 

机构地区：[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

出　　处：《遗传》2014年第6期552-557,共6页Hereditas（Beijing)

基　　金：国家自然科学基金项目(编号:81270343;81100187)资助

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。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 de-tected 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 pre-sented with deletions of both AZFa,b,c and AMEL-Y, indicating sex reversal which was confirmed to be 46,XX by karyo-typing. 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 chromo-some anomalies. It could serve as an effective technique for screening Y-microdeletions, and thus have general application in diagnosis and treatment of male infertility.

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 

分 类 号：R698.2[医药卫生—泌尿科学]