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无精子症ICSI候选男性Y染色体微缺失的多重PCR筛查

Multiplex PCR Screening of Y-chromosome microdeletions in azoospermic ICSI candidate men.

作者信息

Sheikhha Mohammad Hasan, Zaimy Mohammad Ali, Soleimanian Saeede, Kalantar Seyed Mehdi, Rasti Azam, Golzade Maryam, Hoseini Fahraji Hamid

机构信息

Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

Department of Animal Sciences, Agriculture Faculty, Urumia University, Urumia, Iran.

出版信息

Iran J Reprod Med. 2013 Apr;11(4):335-8.

Abstract

BACKGROUND

It has been hypothesized that Y-q microdeletion can account for significant proportion of infertility in men. There are three nonoverlapping regions referred to as the "azoozpermia factors" AZFa, AZFb, and AZFc from proximal to distal part of Y-q. These have been defined as spermatogenesis loci, this region deletions have been shown to be involved in male azoospermic or severe oligoozospermic infertility.

OBJECTIVE

Evaluation the rate of Y-chromosome microdeletions in infertile men.

MATERIALS AND METHODS

In this case-control study, 25 azoospermic infertile men candidate for intracytoplasmic sperm injection (ICSI) were selected as case group. For control group, 25 normoozoospemric men were selected. All cases and controls had normal 46XY karyotype. DNA extraction and molecular analysis were done on blood samples. Multiplex-PCR method was done to identify the presence of microdeletion in AZFa, AZFb or AZFc loci. Eight STS primers that include two controls were selected to determine Y-chromosome microdeletions.

RESULTS

20% (5/25) of all patients have at least one microdeletion in more than one region of AZF loci. Totally 17 microdeletions was observed, one case had deletions in three AZF regions, and 4 cases had deletions in two AZF regions. The rate of deletions was 42% (7/17) for AZFc, 35% (6/17) for AZFa and 23% (4/17) for AZFb.

CONCLUSION

The molecular DNA analysis could help us to know the real cause of infertility and can give good information for good decision for example in men whit microdeletions who want to undertake ICSI procedure the deletions will be passed to their son.

摘要

背景

据推测,Y染色体长臂微缺失可导致相当比例的男性不育。Y染色体长臂从近端到远端有三个不重叠的区域,称为“无精子症因子”AZFa、AZFb和AZFc。这些区域被定义为精子发生位点,已证实该区域的缺失与男性无精子症或严重少精子症不育有关。

目的

评估不育男性Y染色体微缺失的发生率。

材料与方法

在这项病例对照研究中,选择25名接受卵胞浆内单精子注射(ICSI)的无精子症不育男性作为病例组。对照组选择25名精子正常的男性。所有病例和对照的核型均为正常的46XY。对血液样本进行DNA提取和分子分析。采用多重聚合酶链反应(Multiplex-PCR)方法检测AZFa、AZFb或AZFc位点微缺失的存在情况。选择包括两个对照的8个序列特异性引物(STS)来检测Y染色体微缺失。

结果

所有患者中有20%(5/25)在AZF位点的一个以上区域至少有一处微缺失。共观察到17处微缺失,1例在三个AZF区域均有缺失,4例在两个AZF区域有缺失。AZFc区域的缺失率为42%(7/17),AZFa区域为35%(6/17),AZFb区域为23%(4/17)。

结论

分子DNA分析有助于我们了解不育的真正原因,并能为做出正确决策提供有用信息,例如对于有微缺失且想接受ICSI治疗的男性,其微缺失可能会遗传给儿子。

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