罕见46,XY/47,XYY嵌合体型女性患者病因诊断的遗传咨询模式探讨

目的:报道1例46,XY/47,XYY嵌合体女性性反转,探讨其病因诊断路径,提供临床的遗传咨询。方法:临床特征分析和家系调查。采用外周血淋巴细胞核型分析技术进行染色体核型分析,同时基于二代测序技术（NGS）的全基因组拷贝数变异测序（CNV-seq）进行外周血标本拷贝数变异检测及性别决定基因（SRY基因）及AZF基因位点检测。结果:家系调查表明,该患者具有伴X隐性遗传模式;临床上呈现共外显性特征;社会性别为女性,患者的外周血染色体核型结果为46,XY[57%]/47,XYY[13%],染色体拷贝数变异检测示:Arr[hg19]46,XY[70%]/47,XYY[30%],Yp11.31-q11.223×3,dup(6)(q14.1),SRY基因检测阳性,AZF基因所检测位点均未见缺失。结论:细胞遗传学染色体核型分析技术及CNV-seq均可确诊46,XY/47,XYY嵌合体女性性反转综合征,其遗传病因可能为DAX-1基因表达异常。整合了针对46,XY性反转快速诊断路径的遗传咨询模式,供临床参考。     

性反转综合征患者的细胞及分子遗传学分析

目的:探讨检测SRY基因和Y染色体AZF因子在性反转综合征患者临床分子诊断中的应用价值及性反转综合征的机制。方法:运用细胞遗传学核型分析技术和聚合酶链反应(PCR)对7例46,XY女性性反转患者及1例46,XX男性性反转患者进行染色体核型分析、SRY基因扩增和Y染色体AZF因子扩增,SRY阳性的扩增产物进行序列测序。结果:7例46,XY女性性反转中4例SRY和Y染色体AZF因子检测阳性,3例SRY和Y染色体AZF因子检测阴性;1例46,XX男性性反转患者SRY和Y染色体AZF因子检测阴性。4例SRY阳性病例DNA序列分析均未检测到突变。结论:SRY是决定性别和分化的重要基因,Y染色体AZF因子有辅助检测的作用,对性反转综合征患者进行检测有利于明确病因,为其诊断和治疗提供科学依据。性反转综合征可能与其它性别分化相关基因的异常有关,需深入进行细胞、分子遗传学研究。     

特殊特纳综合征1例孕妇的遗传学分析

目的 通过1例特殊特纳综合征孕妇的多种遗传学检测分析,探讨特纳综合征的临床表型及遗传学特点,为该类遗传咨询和临床治疗提供依据.方法 对1例无创DNA产前检测技术(NIPT)检测X染色体异常的孕妇进行遗传学分析,羊水穿刺采用羊水细胞培养及染色体制备技术及荧光原位杂交(FISH)技术分析胎儿染色体;采用染色体核型分析技术及荧光原位杂交技术分析孕妇外周血及口腔脱落细胞.结果 NIPT检测结果显示母体X染色体有缺失;经羊水核型分析,胎儿染色体正常;孕妇外周血染色体核型结果:45,XO;孕妇口腔黏膜脱落细胞FISH结果:47,XXX(65%)/45,XO(23%)/46,XX(12%).结论 将细胞遗传学、分子遗传学以及分子细胞遗传学技术结合使用后能够有效提升特纳综合征诊查的准确性,精准的诊断能够为患有该症患者的遗传咨询以及临床诊治提供参考依据.     

额外小标记染色体嵌合型特纳综合征2例的临床遗传学研究

目的 研究2例携额外小标记染色体(sSMC)特纳氏综合征(TS)患者的临床遗传学情况，探讨Y染色体性别决定区(SRY)在TS患者基因型-表型之间的作用。方法 运用细胞遗传学、荧光原位杂交技术(FISH)对2例携sSMC的TS患者进行检测并对患儿临床资料进行整理分析。结果 经鉴定病例1 sSMC为假等臂双着丝粒Y,核型为45,X[89]/46,X,+mar[11].ish psu dic(Y)(q11.23)(SRY++,DYZ₃++);病例2 sSMC为一罕见环状X,核型为45, X [22]/46,X.ish r(X)(p11.1q13)(DXZ₁+,SRY-),rev ish dim(X)(p11.2p22.3), dim (q21q28)[8]。结论 通过细胞遗传及荧光原位杂交技术可以准确鉴定sSMC来源及组成，SRY阳性与否对携sSMC的TS患者表型及预后具有重要的意义。     

性染色体型性发育异常的遗传学分析及临床意义

目的探讨不同类型性染色体型性发育异常(DSD)患者的遗传学改变及临床意义。方法选取西安交通大学第一附属医院356例性染色体型DSD患者进行G显带染色体分析,对其中18例45,X/46,XY嵌合体者进行Y染色体微缺失检测。结果356例染色体型DSD患者中,克氏综合征(KS)210例,Turner综合征(TS)124例,47,XYY综合征8例,47,XXX综合征8例,45,X/46,XY男性3例,Y染色体结构异常3例。18例45,X/46,XY患者中,3例存在微缺失,其中AZFc位点缺失2例,AZFa+b+c位点缺失1例。结论染色体型DSD以克氏综合征最常见,其次为Turner综合征。对45,X/46,XY男性患者进行Y染色体微缺失检测,有助于明确病因和选择更优的辅助生殖技术方案,实现优生优育。     

原发性闭经的染色体研究

本文对2891例遗传咨询患者中的20例原发性闭经患者,均作外周血淋巴细胞染色体核型分析,结果显示:20例原发性闭经患者中,染色体未见异常者13例(核型46,XX),染色体异常者7例,占35％。其中45,X 2例(属Turner综合征),47,XXX 1例,46,XY 2例,46,XY/46,XX 1例46,XX/46,X,del(X)(q22)1例。本文认为性染色体异常是原发性闭经的主要原因之一,讨论了某些原发性闭经的产前、婴幼儿期诊断及治疗的最佳时期的选择。     

208例身材矮小儿童染色体核型分析

目的:探讨身材矮小儿童细胞遗传学方面的原因。方法:对我院1998年1月~2004年12月208例身材矮小儿童进行外周血淋巴细胞G显带染色体核型分析检查。结果:发现异常核型53例,均为女性,占女性患者的32.12%,其中45,X25例,46,X i(Xq)15例,46,Xdel(Xp)3例,45,XX,-14-21+t(14∶21)3例,45,X/46,XX 2例,45,X/46,Xdel(Xp)1例,45,X/46,X i(Xq)4例。43例男性患儿未检出异常核型,但有11例46,XY,Y=18、1例46,XY,Y=15。结论:染色体异常是儿童身材矮小的重要遗传因素,应引起临床医师的高度重视。     

妊娠中期产前诊断胎儿47,XYY/46,XY异常核型

目的:分析孕中期血清筛查与胎儿染色体异常之间的关系。方法:对1例孕中期血清筛查21-三体风险率为1/260的高危孕妇进行羊水的细胞遗传学检查。结果:羊水细胞染色体核型为47,XYY/46,XY。结论:孕中期妇女血清生化指标的筛查有助于染色体病的检出。     

一例Xq22.3q27.3缺失患者的遗传学分析

目的：对1例无相关特异表型的Turner综合征患者进行遗传学分析,为临床医生识别女性X染色体偏倚失活提供参考。方法：采集1例原发不孕3年的女性及其丈夫和父母的外周静脉血,行染色体核型分析、女方染色体拷贝数变异(CNVs)检测,并进行遗传学分析。结果：原发不孕患者外周血染色体核型为46,X,del(X)(q22.3),其父母及丈夫染色体核型结果均正常。患者CNVs结果提示X染色体q22.3-q27.3缺失40.62Mb,为致病性改变。患者缺失的X染色体失活偏倚比例为98：2,为X染色体极度失活偏倚。结论：临床医生应考虑到女性患者存在X染色体偏倚失活的可能,尽早发现染色体异常患者,以避免生育风险及出生缺陷的发生。     

荧光原位杂交方法检测口腔黏膜脱落细胞在45,X0嵌合体诊断中的意义

目的应用荧光原位杂交(FISH)方法检测45,X0嵌合体患者口腔黏膜脱落细胞,探讨其诊断意义。方法选择3例45,X0嵌合体及可疑嵌合体患者。其中病例1外周血常规染色体核型方法检测,染色体核型为45,X0/46XX,嵌合比为87.0%/13.0%;病例2染色体核型45,X0/46XX,嵌合比为90.0%/10.0%;病例3染色体核型显示单纯45,X0但可疑嵌合体患者。以上患者均采用无菌拭子分别刮取双侧颊黏膜,采集口腔黏膜脱落细胞。应用北京金菩嘉试剂有限公司生产的18/X/Y着丝粒FISH探针进行荧光原位杂交实验。结果口腔黏膜脱落细胞FISH方法检测的3例患者核型分别显示为X0/XX/XXX、X0/XX/XY和X0/XY嵌合体,嵌合比例为77.5%/17.0%/5.5%、61.0%/29.0%/5.0%和88.0%/12.0%;常规外周血淋巴细胞培养后染色体核型分析方法对其中的XXX和XY核型未能准确检出。结论 45,X0患者应用FISH方法检测口腔黏膜脱落细胞,可以无创、快速检出异常核型,对低比例异常嵌合核型可以做出准确诊断。     

性发育异常患者遗传学特点与临床表型分析

目的 探讨染色体核型及SRY基因检测对性发育异常(DSD)的临床诊断价值.方法 回顾性分析332例DSD的染色体核型、SRY基因和临床表型,染色体核型采用外周血淋巴细胞培养G显带分析,SRY基因采用PCR技术扩增.结果 332例DSD患者中检出异常核型100例,检出率为30.12％.在141例社会性别为男性的DSD患者...     

6例性发育异常病残儿的SRY基因分析

目的:对6例性发育异常患儿进行SRY基因检测,为临床诊断提供参考,并对性发育异常机理进行初步探讨。方法:应用PCR扩增及PCR产物直接测序进行SRY基因分析。结果:1例45,XO男性患儿,SRY基因阳性;1例46,XX男性,SRY阴性;1例46,XY女性,SRY基因阳性,对其SRY基因保守区测序结果表明,该区无突变;3例46,XY男性性异常患儿,SRY基因均为阳性。结论:45,XO男性患儿是由于SRY基因易位所至。女性性反转非SRY基因保守区突变所至。男性性反转可能由于常染色体上与性别决定有关的基因突变所至。     

Dysgenetic male pseudohermaphroditism

The authors report a case of a dysgenetic male pseudohermaphroditism with a 45,X/46,XY karyotype in a mosaic form, which was diagnosed in an infant. The one-week-old infant was evaluated because of proximal hypospadias and retention of the right testis. The results of hormonal tests were the followings: serum FSH 5.2 mU/ml; LH: 2.0 mU/ml; testosterone: 144.3 ng/dl; androstendione: 0.42 μg/l; 17-hydroxyprogesterone: 1.12 ng/ml. Chromosomal analysis revealed 45,X/46,XY karyotype. Fluorescent in vitro hybridization showed that 51% of the lymphocytes had the Y chromosome and the SRY gene. Analysis of the SRY showed no deletion in the AZF a,b,c regions. Pelvic magnetic resonance imaging indicated the presence of vagina between the bladder and the rectum, and it showed a mass measuring 15×8 mm in the right inguinal canal as well as an oval gonadal mass with a size of 13×7 mm in the left scrotum. During surgical intervention, performed at the age of one, the right gonad was removed and biopsy of the scrotal testis was performed. Histological examination revealed dysgenetic testis in both sides. The authors emphasize the necessity of cytogenetic and endocrinological investigations of newborns with perineoscrotal hypospadia and bilateral or unilateral maldescent testes immediately after birth. Surgical removal of the dysgenetic testicular tissue located in the abdominal cavity and its histological evaluation provides separation of mixed gonadal dysgenesis, dysgenetic male pseudohermaphroditism, bilateral gonadal dysgenesis and ovotestis in the 45,X/46,XY mosaic cases. An accurate evaluation is necessary for a correct sex assignment and for surgical intervention to prevent neoplastic degeneration of the dysgenetic gonad.     

人类男性Y染色体变异对男性生育力影响的临床分析

目的:探讨Y染色体异常对男性生育能力的影响。方法:对因不育而就诊的4 238例男性患者进行G显带核型分析。结果:共检出Y染色体异常核型550例,占全部被检者12．98％,其中大Y 染色体异常497例;小Y染色体21例;倒位染色体13例;缺失Y染色体12例;47,XYY 4例;45,XO／ 46,XY嵌合体3例;48,XXYY 1例;环状Y染色体1例;Y染色体的平衡易位46,X,t(Y;17)(q12; q25)1例、46,X,t(Y;3)(q11;p11)1例、46,X,t(Y;14)(q12;q22)1例和1例46,X,t(Y;15) (p11;q13)。结论:Y染色体数目与形态结构的异常对男性生育能力有重要影响。     

Fetal sex determination with real time PCR of fetal DNA in maternal plasma

INTRODUCTION: Non-invasive methods using maternal plasma and serum for molecular genetic diagnosis become an important field of interest in prenatal genetic diagnosis. Free fetal DNA in maternal plasma and serum has been shown to be useful for fetal gender determination, and seems to offer a new possibility to perform non-invasive prenatal genetic diagnosis. A possible application is fetal sex determination for couples at risk of X-linked diseases. The aim of this study was to control the reliability and reproducibility of the real-time PCR amplification of the SRY region. MATERIALS AND METHODS: Maternal serum before amniocentesis, and amnionic fluid samples were obtained from 56 pregnant women during the 11th to 22nd weeks of gestation. Real-time PCR analysis of the SRY region was performed in order to determine the fetal sex. Routine karyotyping of cultured amnionic cells was also performed on the samples. Six cases were excluded. RESULTS: In 26 of 50 pregnancies were found male fetuses by cytogenetic analysis. Real time PCR of maternal plasma has been positive for the SRY region in 27 cases. In 47 cases the cytogenetic gender and the real-time PCR result was correlating. In one case of 46,XY karyotype the PCR reaction for SRY region was negative, in two cases of SRY positivity the karyotype was 46,XX. In this study are presented the results of fetal sex determination in maternal plasma using real time PCR method. CONCLUSIONS: The real time PCR detection of fetal DNA in maternal plasma seems to be an easy non-invasive method to determine the fetal sex at this gestational age. Our experience is promising in terms of the specificity and sensitivity of the method.     

Chromosomal disorders in the background of azoospermia

INTRODUCTION: Nowadays more and more couples face the fact that they cannot have babies in spite of many years of trying. The male factor can be identified in about half of these cases. AIM: The aim of this study was to analyse chromosomal alterations in patients with azoospermia. These patients may be candidates for testicular sperm retrieval and intracytoplasmatic sperm injection. MATERIALS AND METHODS: Preoperative evaluation included routine andrological investigation with 2 semen analysis, ultrasound, hormonal and genetic examination. Traditional histological examination and embryological diagnostic of tissue samples was performed. Cryopreservation of retrieved testicular tissue was also done. Between January 2001 and June 2005 73 biopsies were performed in 71 patients for testicular sperm extraction. In order to obtain an exact diagnosis, the traditional cytogenetic methods and fluorescence in situ hybridization analyses were performed in combination with molecular genetic techniques. Patients were offered to participate in the assisted reproduction programme on the base of their genetic results. RESULTS: In this study, the most characteristic cases were numerical deviations, such as 47,XXY (2 cases), mosaic 47,XXY/49,XXXXY (1 case), 47,XYY (1 case) and mosaic 46,XY/45,X (1 case) karyotypes. Non-obstructive azoospermia was diagnosed in 53 patients (79%). CONCLUSIONS: Authors emphasised the importance of cyto- and molecular examinations in cases of genetical disorders. The results provide a chance for patients to be spared from long-drawn moreover psychological burdening examinations. In addition the costs of different clinical intervention could be saved too.     

CHROMOSOMAL ABNORMALITIES IN PATIENTS WITH AZOOSPERMIA IN WESTERN MEXICO

In order to assess the frequency of chromosomal abnormalities in azoospermic males from western Mexico, we carried out a retrospective study in 227 patients. Forty-three (18.9%) cases with an abnormal karyotype were found. The most frequent chromosomal anomaly was 47,XXY, which was identified in 35 subjects (15.4%). In six cases (2.6%), structural aberrations were detected: two Robertsonian translocations [(45,XY,t(13;22)(p11;p11) and (45,XY,t(13;15)(p11;p11)], a Y;autosome translocation [46,XY,der(15)t(Y;15)(q12;p11)], and three mosaics [mos45,X/46,X,idic(Y)(q11)]. In general, these findings are in accordance with those from other surveys and confirm that the XXY aneuploidy is the most frequent chromosomal abnormality in azoospermic individuals.