Yuanyuan Zhang, Qiang Du, Xiaoliang Liu, Wanting Cui, Rong He, Yanyan Zhao
Department of Clinical Genetics, Shengjing Hospital, China Medical University, Shenyang 110004, China.
Center of Reproductive Medicine, Shengjing Hospital, China Medical University, Shenyang 110004, China.
Yi Chuan. 2014 Jun;36(6):552-7. doi: 10.3724/SP.J.1005.2014.0552.
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.
为评估定量荧光聚合酶链反应（QF-PCR）在无精子症因子（AZF）微缺失快速筛查中的应用，采用多重QF-PCR结合毛细管电泳技术，对1218例非梗阻性无精子症或少精子症的不育男性进行AZF区域9个序列标签位点（STS）检测。选择位于性染色体短臂上的AMEL（牙釉蛋白）以及SRY（Y染色体性别决定区）作为内对照。对外周血淋巴细胞吉姆萨染色中期染色体进行核型分析。1218例患者中，105例（8.62%）被鉴定为AZF微缺失。AZFc缺失（67.62%）最为常见，其次是AZFb、c缺失（20.95%）、AZFb缺失（7.62%）和AZFa缺失（3.81%）。5例患者同时出现AZFa、b、c和AMEL-Y缺失，提示性别反转，核型分析证实为46,XX。105例AZF微缺失患者中，16例核型为染色体异常，最常见的是46,XY,Yqh-（75%，12/16）。此外，在总共1218例受检患者中，86例AMEL-X/AMEL-Y比值异常，提示可能存在性染色体异常，其中68例经核型分析证实为性染色体非整倍体。多重QF-PCR能够在一次反应中检测所有标记，也可提示性染色体异常。它可作为筛查Y微缺失的有效技术，因此在男性不育的诊断和治疗中具有广泛应用。
