男性不育患者Y染色体微缺失液态芯片筛查方法的建立及应用

Establishment and application of screening methods on liquid chip for Y chromosome microdeletions in infertile men

目的建立一套全新的基因诊断方法,检测Y染色体无精子因子(azoospermia factor,AZF)区域微缺失,并对Y染色体微缺失与男性不育相关性进行初步探讨。方法按照欧洲男科协会和欧洲分子遗传实验质控网检测指南推荐标准,采用多重PCR-液态芯片技术对648例精子发生障碍的患者和100例合格捐精者进行Y染色体微缺失筛查。结果648例精子发生障碍的患者中,发现62例患者存在Y染色体AZF区域微缺失,对应于5种缺失模式AZFa,AZFb,AZFc,AZFb+c,AZFa+b+c。按区域统计,AZFc区域缺失的频率最高,其次是AZFb,AZFa的检出率最低。无精子症患者中微缺失的发生率为12．31％,严重少精子患者中微缺失发生率为5．43％。100例对照组没有发现任何缺失,两组比较,差异显著(P<0．001)。结论男性不育与Y染色体微缺失密切相关,本研究建立的多重PCR方法-液态芯片技术平台,用于男性不育患者的YqAZF区域筛查,结果可靠、快捷、重复性好、通量高。

Objective To establish a new method, suitable for clinical gene diagnosis of Y chromosome microdeletions in azoospermia factor （AZF） regions. Explore the possible associations with Y chromosome microdeletions and male infertility. Methods According to the guidelines made by European Academy of Andrology （EAA） and European Molecular Genetics Quality Network （EMQN）, multiple PCR method and liquid gene chip multi-analyte suspension array technology were used to detect the microdeletions of 648 infertile males and 100 normal men. Results 62 cases exist micordeletions among 648 infertile males.There were five deletion models on Y chromosome： AZFa, AZFb, AZFc, AZFa＋b, AZFb＋c, AZFa＋b＋c. According to the study the most common deletions was first occurred in AZFc region, then followed by AZFb region, the least was occurred in AZFa region. About 12.31% of azoospermia cases and 5.43% of oligozoospermia cases with microdeletions were detected in infertile males. No deletions were found among normal men, the difference between the two groups was significant（P〈0.001）. Conclusion There are close relationship with Y chromosome microdeletions and male infertility. The PCR method and liquid chip platform can provide an accurate, rapid, good reproducibile diagnostic and high-throughput method for the detection of Yq.