伴及不伴精索静脉曲张的无精子、严重少精子症患者Y染色体微缺失对比研究

目的:比较精索静脉曲张(VC)无精子症和严重少精子症与不伴VC无精子症和严重少精子症患者Y染色体微缺失发生率,探讨他们不育的内在原因。方法:A组为VC无精子症和严重少精子症的患者137例,其中无精子症70例(A1组),严重少精子症67例(A2组);B组为不伴有VC的特发性无精子症和严重少精子症患者135例,其中无精子症69例(B1组),严重少精子症66例(B2组)。C组(对照组)为30例正常生育男性。采用多重PCR技术对受试者进行Y染色体微缺失检测。结果:1 A组137例中有23例检测到Y染色体微缺失,缺失率16.8%。B组135例中有23例检测到Y染色体微缺失,缺失率17.0%;C组未检测到Y染色体微缺失;2 A1组、A2组、B1组和B2组Y染色体微缺失率分别为为22.9%、10.4%、20.3%和13.6%;3严重少精子症A2组和B2组共133例中16例检测出Y染色体微缺失,发生率为12.0%;4A组与B组比较,差异无统计学意义(P0.05)。结论:Y染色体微缺失发生率在伴有及不伴有精索静脉曲张的无精子、严重少精子症患者中无显著差异,Y染色体微缺失是精索静脉曲张伴有的无精子、严重少精子症病因之一。     

特发性无精子症和严重少精子症患者Y染色体基因微缺失研究

目的:研究Y染色体基因微缺失与特发性无精子症和严重少精子症的关系,及探讨Y染色体基因微缺失的位点、缺失率有无民族间的差异性.方法:应用多重实时荧光定量聚合酶链反应(PCR)法,对40例汉族及维吾尔族特发性无精子和严重少精子症患者进行Y染色体Azoospermia Factor(AZF)因子多位点的微缺失检测.结果:23例特发性无精子患者中,3例发生AZF因子缺失,缺失率为13.04%;17例严重少精子症患者中,2例发生AZF因子缺失,缺失率为11.76%.结论:Y染色体AZF因子微缺失的范围和位置对于胞质内体外受精治疗男性不育具有重要意义,但Y染色体AZF因子的缺失的位点及缺失率有无民族间的差异性,值得进一步研究.     

陕西地区不明原因无精子症和少精子症患者Y染色体长臂微缺失分析

目的: 评估陕西地区不明原因无精子症和少精子症不育男性患者Y染色体长臂微缺失的频率,探讨精子密度与Y染色体微缺失发生率的相关性。　方法: 以Y染色体特异性无精子症因子区STS AZFa、AZFb、AZFc和SRY4个基因 5个片段设计引物,采用PCR方法对 64例无精子症和少精子症患者以及 20例正常生育男性进行微缺失检测,并比较不同精子密度患者Y染色体微缺失的发生率。　结果: 20例精子密度正常的生育男性未检出Y染色体微缺失,而 64例特发性无精子症 /少精子症患者AZFc区的缺失率为17. 2% (11 /64),AZFc和AZFb联合缺失 1例,未发现AZFa区缺失,SRY基因均为阳性。其中无精子症组缺失率为21. 43% ( 3 /14 );精子密度 <1×106 /ml组,缺失率为 20. 0% (2 /10);精子密度 (1 ~5)×106 /ml组缺失率为17. 9% (5 /28);精子密度 (5 ~10 )×106 /ml组缺失率为8. 3% (1 /12)。各组缺失率经卡方检验差异有显著性 (χ2 =70. 144,P<0. 005 )。　结论: 无精子症和少精子症不育患者Y染色体AZFc缺失率明显较高,PCR扩增AZF基因是诊断Y染色体微缺失的简单方法。     

成年多囊肾病与男性不育（附5例分析）

目的 观察多囊肾病(PKD)患者泌尿生殖系统改变及其对男性生育力的影响.方法 对3318例男性不育患者进行B超探查精囊腺、前列腺、睾丸、附睾、精索、肾脏和肝脏等,精液常规分析,232例正常生育成年男性作为对照组.多重PCR检测Y染色体无精子因子(azoospermia factor,AZF)微缺失.结果 3318例男性不育患者中PKD5例(0.15%,5/3318).发现PKD患者一家系7例.5例PKD患者(附睾囊肿5例,精囊囊肿3例),其中1例为无精子症,2例严重少精子症(精子密度＜5×106/ml),2例少精子、死精子症(精子密度＜20×106/ml而＞5×106/ml,100%死精子),检测到Y染色体AZF缺失1例.结论 PKD合并生殖系统的囊性化阻塞可能是造成精子运输障碍引起男性不育的主要原因.     

Y染色体微缺失与无精子症少精子症关系的研究

目的 探讨Y染色体微缺失与无精子症、少精子症的关系.方法 应用多重聚合酶链反应技术(PCR)对127例无精子症(80例)和严重少精子症(47例)的不育患者及60例正常生育男性进行Y染色体AZF基因、DAZ外显子检测.结果 无精子和严重少精子患者Y染色体微缺失7例,缺失率5.51%.其中AZFc缺失2例,DAZ外显子缺失5例.少精子症组缺失率8.51%,无精子症组缺失率3.75%,小睾丸组的缺失率6.54%,正常睾丸组缺失率4.94%,正常生育男性AZF基因和DAZ外显子均未检测到缺失.结论 (1)AZF因子、DAZ外显子微缺失可导致无精子症、严重少精子症:(2)绝大部分无精子、严重少精子患者Y染色体AZF因子、DAZ外显子并没有微缺失,有必要再去寻找新的精子发生基因.     

多重聚合酶链反应检测Y染色体无精子因子微缺失

目的:研究原发性无精、严重少精症与Y染色体无精子因子(AZF)微缺失之间的关系.方法:采用多重聚合酶链反应技术对103例原发无精子症、72例原发严重少精症患者及60例正常生育男性进行AZFa、AZFb、AZFc 3个区域微缺失分析.结果:60例正常生育男性未发现Y染色体AZF区域微缺失,175例生精障碍患者中发现AZF微缺失19例,总缺失率为10.9%.其中11例无精症患者和4例少精症患者的缺失发生在AZFc区域,缺失率为8.6%;1例无精症患者和2例少精症患者发生AZFb、AZFc双重缺失,缺失率为1.7%;1例无精症患者发生AZFa、b、c 3个区域同时微缺失,缺失率0.6%.生精障碍组与正常生育男性组比较Y染色体AZF区域微缺失率差异有统计学意义(P<0.01).结论:Y染色体AZF区域微缺失是引起男性无精、少精子症的重要原因之一.采用多重聚合酶链反应技术对原发无精、少精子症患者在单精子注射(ICSI)之前进行微缺失筛查是必要的.     

无精子症和严重少精子症134例遗传学分析

目的：研究无精子症和严重少精子症患者染色体畸变及Y染色体(Yql1区)无精子症因子(azoospermic factor，AZF)缺失情况，建立Y染色体微缺失的临床筛查方法。方法：对134例患者(无精子症97例，严重少精子症37例)经染色体核型分析及AZF、区三个位点8对引物PCR扩增，检测染色体畸变和Y染色体微缺失率。结果：134例中染色体核型异常9例，占6．72％。AZF缺失18例，缺失率为13．43％。无精子症和严重少精子症AZF、缺失率分别为14．43％、10．81％。结论：染色体畸变和Y染色体微缺失是导致无精子症和严重少精子症的主要原因之一。无精子症缺失率高于严重少精子症患者。AZF区三个位点8对引物PCR扩增可作为Y染色体微缺失的临床筛查方法。     

新疆地区汉族维吾尔族不育症患者Y染色体长臂微缺失分析

目的评估新疆地区汉族、维吾尔族不明原因无精子症和严重少精子症男性患者Y染色体长臂微缺失的频率,探讨不同民族间Y染色体长臂微缺失发生率的差异。方法以Y染色体无精子因子(AZF)区STS- AZFa、AZFb、AZFc和AZFd 4个基因8片段设计引物,采用聚合酶链反应(PCR)方法对123例(汉族61例,维吾尔族62例)无精子症和少精予症不育男性患者进行Y染色体微缺失检测,并比较不同民族的患者Y染色体微缺失发生率的差异。结果61例汉族患者中有27例(44.26%)存在Y染色体微缺失,62例维吾尔族患者检出13例(20.97%)存在Y染色体微缺失,在所有被检出有Y染色体长臂微缺失的患者中AZF区联合缺失23例(58%)。汉族患者与维吾尔族患者Y染色体微缺失率及AZF多位点联合缺失发生率差异有统计学意义(P<0.05)。结论无精子症和严重少精子症不育男性患者中Y染色体长臂微缺失发生率及AZF多位点联合缺失发生率存在民族差异,PCR检测AZF基因是诊断Y染色体长臂微缺失的较好的方法。     

特发性无精子症和严重少精子症患者无精子因子的检测及其意义

目的：探讨引起特发性无精子和严重少精子造成男性不育的遗传学原因和检测无精子因子(AZF)的临床意义。方法：对50例特发性男性不育患者(不育组)和50例正常生育者(对照组)的外周血标本．提取基因组DNA,通过多重聚合酶链反应检测Y染色体AZt?微缺失。结果：对照组均可见SRY、SY84、SY86、YRRM1(RBM1)和SY254(DAZ)扩增带。不育组6例(无精子症4例．严重少精子症2例)可见SRY扩增带．但未见SY254扩增带,其中2例同时未见YRRM。扩增带;1例仅未见YRRM。扩增带。结论：Y染色体AZF微缺失是引起无精子和严重少精子并造成男性不育的重要原因之一;AZF微缺失检测对男性不育症患者进行遗传学诊断与筛查有一定意义。     

男性生精障碍的细胞遗传学和分子遗传学检测

目的从遗传学角度分析男性生精障碍的病因,为临床提供治疗和遗传咨询的依据。方法对91例无精子症患者和42例严重少精子症患者,采用外周血染色体核型分析和Y染色体AZF区域微缺失联合检测。结果91例原发性无精子症患者中,染色体数量异常者16例,占总数17.5%;染色体平衡易位5例,占总数5%;10例AZF区域STS位点缺失,占总数11%;二项检测异常发生率为34%。42例严重少精子症患者检出染色体平衡易位4例,占总数9.5%;AZF区域STS位点缺失5例,占总数11.9%,二项检测异常发生率为21.4%。结论染色体核型分析和Y染色体微缺失是男性生精障碍重要的遗传检测指标。     

原因不明性无精症和少精症Y染色体微缺失的筛查分析

目的 探讨原因不明性无精症和少精症不男性与Y染色体微缺失的关系。方法 应用多重PCR技术,对38例原因不明性无精症和少精症者（无精症11例、严重少精症9例、少精症18例）基因组DNA进行Y染色体连锁的18个序列标记位点缺失检测。结果 38例中发现Y染色体微缺失6例,缺失率为16％,其中无精症2例,严重少精症1例,少精症3例。缺失形式前两者为AZFd(DYS 237) AZFc(DAZ DYS240),后者为AZFd(DYS237)。结论 Y染色体微缺失是原因不明性无精症和少精症的重要原因之一。采用多重PCR技术进行缺失检测,是一种非常有效的方法。     

Chromosomal abnormalities and y chromosome microdeletions in infertile men with varicocele and idiopathic infertility of South Indian origin

Various factors cause spermatogenesis arrest in men and, in a large number of cases, the underlying reason still remains unknown. Little attention is paid to determining the genetic defects of varicocele-related infertility. The objective of our present study was to investigate the chromosomal abnormalities and Y chromosome microdeletions in infertile men of South Indian origin with varicocele and idiopathic infertility. Metaphase chromosomes of 251 infertile men with varicocele and unexplained infertility were analyzed using Giemsa-Trypsin-Giemsa (GTG) banding and fluorescence in situ hybridization (FISH). The microdeletions in 6 genes and 18 sequence-tagged-sites (STS) in the Yq region were screened using polymerase chain reaction (PCR) techniques. Out of 251 infertile men, 57 (22.7%) men were with varicocele, of which 8.77% were azoospermic, 26.31% were severely oligozoospermic, 21.05% were mildly oligozoospermic, and 43.85% were oligoasthenoteratozoospermic (OAT), and 194 (77.29%), with idiopathic infertility, of which 51% were azoospermic, 13.40% were severely oligozoospermic, 19.07% were mildly oligozoospermic, and 16.4% were with OAT. Genetic defects were observed in 38 (15.13%) infertile individuals, including 14 (24.56%) men with varicocele and 24 (12.37%) men with idiopathic infertility. The frequencies of chromosomal defects in varicocele and idiopathic infertility were 19.3% and 8.76%, respectively, whereas Y chromosome microdeletions were 5.26% and 3.60%, respectively. Overall rate of incidence of chromosomal anomalies and microdeletions in 251 infertile men were 11.5% and 3.98%, respectively, indicating a very significant higher association of genetic defects with varicocele than idiopathic male infertility. Our data also demonstrate that, among infertile men with varicocele, severely oligozoospermic and OAT men with varicocele have higher incidences of genetic defects than mildly oligozoospermic and azoospermic men.     

Y chromosome microdeletions and varicocele as aetiological factors of male infertility: A cross‐sectional study

The pathogenic mechanisms by which varicocele disrupt spermatogenesis are not clearly understood. Over 30% of male infertility cases resulting from spermatogenic problems are associated with genetic abnormalities, and Y chromosome microdeletions are the second most frequent genetic cause. Here, we aimed to evaluate the frequency of Y chromosome microdeletion in infertile men with varicocele. A cross‐sectional study comprising 51 infertile men with varicocele presenting spermatogenesis failures was performed. Y chromosome microdeletion research was made using polymerase chain reaction. Of the 51 men with infertility and varicocele, 35.3% (18/51) had nonobstructive azoospermia and 64.7% had severe oligozoospermia. Y chromosome microdeletion was found in two cases (3.9%): one patient had nonobstructive azoospermia and complete microdeletion of the AZFb and AZFc regions, and another patient had severe oligozoospermia and complete microdeletion of the AZFc region. Although in recent years, a genetic aetiology related to Y chromosome microdeletions has become a major cause of infertility in males with spermatogenesis failures, in this study, the varicocele was the clinical cause of seminal abnormalities that could lead to infertility, suggesting that both varicocele and Y chromosome microdeletion aetiologies can present, alone or combined, as factors of male infertility.     

Screening of Y chromosome microdeletions in Tunisian infertile men

The aim of this study was to establish the prevalence of Y chromosomal microdeletions in infertile Tunisian men. Three groups of infertile men, 65 normospermic, 53 oligozoospermic and 45 azoospermic, were tested for Yq microdeletions detection by multiplex polymerase chain reaction (PCR) using specific Y chromosome AZF regions tagged site markers (STS). One group of 13 healthy men was used as the control group. Six STS were tested (2 in each AZF region). The general prevalence of AZF microdeletions was 16%; in azoospermia and severe oligospermia groups, it was higher (29% and 30.5%, respectively). Significant differences were found with moderate oligospermic and normospermic groups (p < 0,05). AZFc microdeletions were the most frequent, and 55% of AZFc deleted patients were oligospermic. No deletions were detected in the control group. These results add to the growing literature data, showing that microdeletions of the Y chromosome is an important cause of severe spermatogenetic defect and confirm that deletion in AZFc region is the most common and is compatible with residual spermatogenesis.     

Screening for microdeletions on the long arm of chromosome Y in 53 infertile men

About 30% of couple infertilities are of male origin. They appear in some cases de novo and are considered idiopathic. The aim of our work was to evaluate, in these cases, the prevalence of microdeletions of the long arm of chromosome Y, within the AZF a, b and c regions using molecular biology techniques. Men with azoospermia or oligozoospermia resulting from hereditary, endocrine or obstructive causes, or with a constitutional cytogenetic abnormality were excluded. Fifty-three infertile men with azoospermia or oligozoospermia, as determined by a spermiogram, were studied. Of these, 34 were idiopathic and 7 exhibited a past history of genital infection or biological abnormalities, suggesting partial obstruction of the genito-urinary tract. A further 8 men had a varicocele and 11 cases with a history of cryptorchidism were also studied. Peripheral blood DNA was extracted from each patient, then amplified by multiplex PCR with STS genomic markers from the three Y chromosome AZF zones. PCR products were then analysed on agarose gels. In view of the difficulty of confirming the absence of a signal in molecular biology, each case suspected of having a deletion was checked by multiplex PCR through coamplification with the SRY marker. Five men with microdeletions of the long arm of the Y chromosome were diagnosed among the 53 patients. All of them included the AZFc zone and the intragenic DAZ gene markers. Furthermore, a larger Y chromosome deletion encompassing the 3 AZF zones was diagnosed, and confirmed by cytogenetic analysis. All Y chromosome microdeletions were observed in the 34 truly idiopathic azoospermia/oligozoospermia cases, corresponding to a proportion of 14.7% (or 9.4% considering the whole population of 53 infertile men). The relatively high proportion of microdeletions found in our series suggests the need for strict patient selection to avoid unnecessary screening for long arm Y chromosome microdeletions.     

Cytogenetic and molecular study of 370 infertile men in South India highlighting the importance of copy number variations by multiplex ligation-dependent probe amplification

Male infertility is a common and severe problem affecting 7% of population. The main objective of this study is to identify the chromosomal abnormalities, Y microdeletions in infertile men and also to access the frequency of abnormal sperm count. Based on the sperm count and viability, the infertile men were grouped as Azoospermia, Asthenospermia, Oligospermia and the remaining as Idiopathic infertility. A total of 370 infertile men and 60 normal control men were recruited. Chromosomal abnormalities were identified in 3 men (3/370). The prevalence of Y microdeletions in the infertile group is 8/370 in the Azoospermia factor (AZF) region with four AZFc deletion/duplication, two AZFa deletion, one AZF b & AZFc deletion and one case of total AZF a, AZFb & AZFc deletion. However, only five cases of Y microdeletions were identified by Multiplex PCR but an additional three cases by MLPA (Multiplex ligation-dependent probe amplification). Fluorescence in situ hybridisation also confirmed the deletions. Here, we performed MLPA post-multiplex PCR, and our study revealed good yield of the Y microdeletion identification. The partial duplications which are difficult to be identified can now be easily identified by MLPA, and hence, we recommend MLPA as the choice of investigation compared to multiplex PCR for infertile men.     

Histology and sperm retrieval among men with Y chromosome microdeletions

In this review of Y chromosome microdeletions, azoospermia factor (AZF) deletion subtypes, histological features and microTESE sperm retrieval rates are summarized after a systematic literature review. PubMed was searched and papers were identified using Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Approximately half of infertile couples have a male factor contributing to their infertility. One of the most common genetic etiologies are Y chromosome microdeletions. Men with Y chromosome microdeletions may have rare sperm available in the ejaculate or undergo surgical sperm retrieval and subsequent intracytoplasmic sperm injection to produce offspring. Azoospermia or severe oligozoospermia are the most common semen analysis findings found in men with Y chromosome microdeletions, associated with impaired spermatogenesis. Men with complete deletions of azoospermia factor a, b, or a combination of any loci have severely impaired spermatogenesis and are nearly always azoospermic with no sperm retrievable from the testis. Deletions of the azoospermia factor c or d often have sperm production and the highest likelihood of a successful sperm retrieval. In men with AZFc deletions, histologically, 46% of men demonstrate Sertoli cell only syndrome on biopsy, whereas 38.2% have maturation arrest and 15.7% have hypospermatogenesis. The microTESE sperm retrieval rates in AZFc-deleted men range from 13-100% based on the 32 studies analyzed, with a mean sperm retrieval rate of 47%.