原发闭经表型的细胞遗传学病因研究

目的:探讨原发闭经表型的细胞遗传学病因。方法:运用常规的染色体核型分析技术及FISH、C-带和Q-带技术,分析25例原发闭经表型患者的染色体核型。结果:25例原发闭经表型病例中,18例核型异常,占72.0%。异常核型病例分别为:45,X 8例、46,XY 2例,等臂X 1例、嵌合核型4例(分别为45,X细胞系与不同的结构异常性染色体细胞系的嵌合体)、X长臂末端缺失1例、X-常染色体平衡易位2例。除这18例异常核型患者外,尚有7例为先天性无子宫、无阴道的原发闭经患者未检出核型异常。结论:X染色体数目单体、X结构异常、X染色体数目单体细胞系与结构异常性染色体细胞系的嵌合体核型、XY性发育异常(DSD)、X-常染色体平衡易位都可导致原发闭经表型。常规G-显带技术与C-带、Q-带技术及FISH技术相互结合,有助于进一步准确诊断核型。     

应用荧光原位杂交技术检测卵巢早衰患者X染色体嵌合型

目的　探讨卵巢早衰与低百分率 45 ,X/ 46,XX嵌合核型的关系 ,了解荧光原位杂交(FISH)技术检测染色体嵌合核型的敏感性和特异性。方法　取 18例卵巢早衰患者和 9例正常妇女(对照 )的外周血进行培养 ,行常规G显带和用X染色体记数探针行FISH ,对每例计数 60 0～ 70 0个细胞的荧光信号。结果　 18例卵巢早衰患者的常规染色体分析核型为 46,XX。卵巢早衰患者的 45 ,X/46,XX染色体嵌合型比率为 7.6% ,显著高于对照者的 2 .2 % ,两者比较 ,差异有极显著性 (P <0 .0 1)。结论　某些卵巢早衰患者的致病机理可能与 45 ,X/ 46,XX嵌合核型比例增高 ,X染色体数量不足有关。对此类低百分率染色体嵌合核型 ,FISH技术在敏感性和特异性上优于常规核型检查     

Y染色体长臂缺失及不分离不育男性1例报道

目的:报道1例Y染色体长臂缺失合并不分离的男性无精子症患者。方法:常规染色体核型分析,荧光原位杂交以确定核型。PCR-STSs检测以确定Y染色体断裂点,并行睾丸活检。结果:细胞遗传学和FISH证实患者为嵌合体,核型为45,X/46,X,del(Y)/47,X,del(Y)del(Y)。分别占27%,68%,5%。C带显示患者Yq12全部丢失。PCR-STSs检测AZFa存在,AZFb和AZFc区域全部丢失,断裂点位于sY88和sY95之间及sY88以下。睾丸病理显示精曲小管中只有支持细胞,没有生精细胞。未见卵巢组织。结论:患者无精子症、睾丸体积小与病理结果一致,其原因是由于Yq11.2的缺失。     

国产13、18、21、X、Y五色探针在产前遗传病诊断中的应用价值

目的 探讨国产21,13,18,X,Y五色探针诊断胎儿最常见染色体疾病的应用价值。方法 采集101例孕周l4~22周孕妇羊水标本,应用国产检测试剂盒21,13,18,X,Y探针进行羊水间期细胞FISH检测,其结果与羊水细胞学染色体培养结果进行对照,计算其灵敏度、特异性、Kappa值等。结果 101例FISH检测全部成功,未见异常99例,发现2例异常核型:21三体嵌合体(47XX,+21/46XX),18三体(47,XY,+18)。并与细胞学染色体培养结果进行对照,所得结果均一致,符合率,灵敏度,特异性,kappa值,均为100%。结论国产五色荧光探针与羊水间期细胞杂交可快速诊断胎儿21,13,18,X和Y染色体数目异常。结果可疑者,要进行常规染色体核型分析。     

荧光原位杂交技术快速产前诊断胎儿血细胞非整倍体的价值

目的 探讨荧光原位杂交（FISH）技术在产前诊断脐血细胞非整倍体中的应用价值：方法 2004-06—2005-03，对广州市妇婴医院114例孕18～38W有产前诊断指征的孕妇进行脐血穿刺。采用X／Y染色体着丝粒探针和21q22．13-q22．2特异性探针对脐血细胞进行间期FISH检测，然后在荧光显微镜下观察，用Leica染色体核型自动分析仪QFISH软件进行图像的摄取和处理。同时所有脐血标本进行细胞培养，常规染色体G显带核型分析作为对照。结果 114例脐血标本都有FISH检测结果，107例具有正常核型染色体数目，异常7例，其中4例为唐氏综合征（3例为典型唐氏综合征，1例为嵌舍体），3例为性染色体数目异常。结论 FISH技术用于产前诊断脐血常见染色体数目异常，具有简便、快速、特异性强等优点，能为临床诊断提供依据。     

516例无精症Y染色体微缺失及细胞遗传学分析

目的 探讨Y染色体微缺失和细胞遗传学分析在无精症中的相关性.方法 对2015年3月至2019年12月来广东省妇幼保健院就诊的诊断为无精症的患者进行外周血染色体G显带核型分析,运用多重定量荧光聚合酶链反应(quantitative fluorescent PCR,QF-PCR)技术检测Y染色体上的无精因子(azoospe...     

应用FISH技术分析一例平衡易位t(1;16)伴反复流产

目的: 应用FISH技术分析报道一例伴有t(1;16)的反复流产(RSA)者。方法: 用Biotin-11 -dUTP标记的1号染色体长臂末端探针(1qter)和16号染色体着丝粒特异性α卫星DNA探针(16cen)以及CY3标记的1号染色体着丝粒特异性α卫星DNA探针(1cen)对1例常规G显带核型分析为46,XY,t(1;16)(q21; q13)的反复流产患者进行双色荧光原位杂交研究。 结果: 双色FISH显示该患者t(1;16)易位所致的衍生染色体着丝粒均含有红绿两个信号融合而成。 结论: 双色FISH证实衍生的染色体着丝粒是由1号和16号染色体着丝粒共同组成,该患者最后的核型确诊为46,XY, t(1;16) (p10; q10)。FISH技术是检测染色体结构异常的一种良好的辅助手段。     

FISH技术在检测自然流产绒毛组织非整倍体异常中的应用研究

目的:探讨运用荧光原位杂交(FISH)检测自然流产绒毛组织的临床价值,评价它与传统经典的核型分析方法的关系。方法:对157例孕早期自然流产的绒毛组织进行FISH检测,均采用16、22、13、21、18、X、Y号染色体荧光探针检测,判断染色体非整倍体异常情况。同时进行绒毛细胞培养染色体核型分析,作为对照诊断标准。结果:核型分析成功率为48.4%,FISH检测成功率为100%。核型分析成功的76例样本中,64例结果与核型分析结果相一致,以细胞遗传学作为诊断标准,诊断的符合率为84.2%。结论:FISH技术与传统的绒毛细胞培养染色体核型分析相比,过程迅速,方法简单,提高了诊断的成功率,但无法完全取代传统的染色体核型分析,应两者结合应用于临床。     

Turner综合征患者标记染色体的鉴定与分析

目的 分析11例携带标记染色体的Turner综合征患者的核型,研究这类染色体的表型效应。方法 选择11例具Turner综合征表型的患者,常规核型分析均显示为携带标记染色体的嵌合体,其中6例标记染色体呈环状。患者G带核型表示为mos．45,X／46,X,＋mar或者mos．45,X／46,X,＋r．以X／Y着丝粒探针,应用荧光原位杂交（FISH）技术分析这些标记染色体起源,对其中2例较大的环状染色体,结合反向染色体涂染确定断裂位点,比较不同断裂位点的标记染色体的遗传学效应。结果11例患者所携带的标记染色体均为环状染色体,r（X）的断裂位点分别位于Xp22、Xq22、Xq24、Xq26等。结论 Turner综合征患者的标记染色体主要来源于X染色体,且表现为r（X）形式。r（X）均以嵌合型的形式存在。     

FISH技术在流产组织检查及产前诊断中的应用研究

目的探讨FISH技术诊断流产组织、未培养羊水细胞染色体异常的临床应用价值。方法采用着丝粒、专一序列探针对流产绒毛组织、胎儿组织及产前诊断孕妇的羊水细胞进行FISH检测;并与常规细胞遗传学方法进行对比。结果 7例流产绒毛组织中FISH技术检测出5例染色体异常,分别是3例nuc ish 22q11×3,1例nuc ish 16q22×3,1例三倍体;2例流产胎儿组织中FISH技术未发现异常。孕中期产前诊断的156例孕妇,常规细胞遗传学方法发现3例胎儿染色体异常,分别为47,XX,+21 1例,46,XX,dup(?q21q22)1例,47,XXY 1例;FISH技术与常规细胞遗传学方法检测结果基本一致,在病例46,XX,dup(?q21q22)中,G显带400带水平进行染色体核型分析不能确定其21号染色体重复的确切区带,而FISH结果为nuc ish 21q22×3,证实探针杂交区域有重复。另1例孕晚期羊水FISH检测结果未见异常。结论 FISH技术在流产胚胎染色体检查及产前诊断中具有检测快速,操作简便,标本采集及取材时间不受限等优势,目前FISH技术也存在探针位点有限及无法检测染色体结构异常等问题。     

High-level mosaicism for 45,X in 45,X/46,X,idic(Y)(q11.2) at amniocentesis in a pregnancy with a favorable outcome and postnatal progressive decrease of the 45,X cell line

ObjectiveWe present prenatal diagnosis of high-level mosaicism for 45,X in 45,X/46,X,idic(Y)(q11.2) at amniocentesis in a pregnancy with a favorable outcome and postnatal progressive decrease of the 45,X cell line.Case reportA 36-year-old, gravida 4, para 3, woman underwent amniocentesis at 17 weeks of gestation because of advanced maternal age. Amniocentesis revealed a karyotype of 45,X[22]/46,X,idic(Y)(q11.2)[4]. Prenatal ultrasound was unremarkable, and the fetus had normal male external genitalia. Repeat amniocentesis was performed at 20 weeks of gestation, and the second amniocentesis revealed a karyotype of 45,X[24]/46,X,idic(Y)(q11.2)[3]. Simultaneous interphase fluorescence in situ hybridization (FISH) analysis on uncultured amniocytes revealed that 60% (62/103 cells) were Y-deleted cells. After genetic counseling, the parents decided to continue the pregnancy, and a 3020-g male baby was delivered with a body length of 52 cm, normal male genital organs and no phenotypic abnormalities. The karyotypes of cord blood, umbilical cord and placenta were 45,X[20]/46,X,idic(Y)(q11.2)[20], 45,X[31]/46,X,idic(Y)(q11.2)[9] and 45,X[40], respectively. At age one month, FISH analysis on urinary cells and buccal mucosal cells revealed 11.5% (7/61 cells) and 13.6% (16/118 cells), respectively for mosaicism for the Y-deleted cells. At age five month, the karyotype of peripheral blood was 45,X[9]/46,X,idic(Y)(q11.2)[31]. FISH analysis on buccal mucosal cells showed no abnormal Y-deleted cell (0/101 cells). At age 11 month, the karyotype of peripheral blood was 45,X[5]/46,X,idic(Y)(q11.2)[35]. FISH analysis on 102 buccal mucosal cells showed no abnormal signals. The infant was doing well with normal physical and psychomotor development.ConclusionHigh-level mosaicism for 45,X in 45,X/46,X,idic(Y)(q11.2) at amniocentesis can be associated with a favorable outcome and progressive decrease of the 45,X cell line.     

Prenatal and postnatal characterization of Y chromosome structural anomalies by molecular cytogenetic analysis

We describe three cases in which we used fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR) and comparative genomic hybridization (CGH) to characterize Y chromosome structural anomalies, unidentifiable by conventional G-banding. Case 1 was a 46,X,+mar karyotype; FISH analysis revealed an entire marker chromosome highlighted after hybridization with the Y chromosome painting probe. The PCR study showed the presence of Y chromosome markers AMG and SY620 and the absence of SY143, SY254 and SY147. CGH results confirmed the loss of Yq11.2-qter. These results indicated the presence of a deletion: del(Y)(q11.2). Case 2 was a 45,X [14]/46,XY[86] karyotype with a very small Y chromosome. The PCR study showed the presence of Y chromosome markers SY620 and AMG, and the absence of SY143, SY254 and SY147. CGH results showed gain of Yq11.2-pter and loss of Yq11.2-q12. These results show the presence of a Yp isodicentric: idic(Y)(q11.2). Case 3 was a 45,X,inv(9)(p11q12)[30]/46,X,idic(Y)(p11.3?),inv(9)(p11q12)[70] karyotype. The FISH signal covered all the abnormal Y chromosome using a Y chromosome paint. The PCR study showed the presence of Y chromosome markers AMG, SY620, SY143, SY254 and SY147. CGH only showed gain of Yq11.2-qter. These results support the presence of an unbalanced (Y;Y) translocation. Our results show that the combined use of molecular and classical cytogenetic methods in clinical diagnosis may allow a better delineation of the chromosome regions implicated in specific clinical disorders.     

Turner syndrome in a girl with marker chromosome in karyotype]

OBJECTIVES: There are suggestion, that Turner syndrome (TS) patients with mosaic karyotype for a Y-DNA-containing cell line are at risk of Y-induced gonadoblastoma. The TS patients in whom some or all cells contain a marker chromosome of unknown origin and those in whom there is clitoromegaly or other evident virillisation should be tested by FISH or PCR techniques. DESIGN: The aim of our study to present a TS girl with mosaic karyotype and marker chromosome, which origin from X chromosome was detected by FISH method. MATERIAL AND METHODS: 5-years old girl in whom TS was established. Clinical analysis included the full dysmorphic and clinical phenotype of TS. Chromosome analysis was performed on peripheral blood samples using routine cytogenetic methods and FISH technique. RESULTS: Clinical examination of girl showed many typical signs of TS besides of normal weight and length at birth and not typical for TS patients heart defect. First routine chromosome analysis, at age of 6 month, showed only 45,X cell line, Second study revealed mosaic karyotype with marker chromosome. FISH analysis for interphase nuclei and metaphase chromosomes using X centromere probe explained origin of marker from X chromosome. The karyotype was 45,X[155]/46,X,+mar[8].fish mar(X)(DXZ1+). CONCLUSION: Presence of marker chromosome in karyotype of patient with TS may modify their phenotype and it is a indication for molecular examination by FISH technique.     

Prenatal investigation of a 45,X/46,X,r(?) karyotype in amniocytes using fluorescence in situ hybridization with an X-centromeric probe.

The karyotype of cultured amniotic fluid cells obtained on the indication of advanced maternal age was shown to be a mosaic 45,X/46,X,r(?). The small size and banding pattern made it difficult to determine whether the ring was derived from and X or a Y chromosome, or even from an autosome. By using an X-centromeric probe and fluorescence in situ hybridization (FISH), we demonstrated the ring to have an X centromere. Thus, a more complete genetic counselling was possible. This confirms the usefulness of FISH in identifying and characterizing this and other chromosome rearrangements in prenatal diagnosis.     

Mosaicism for Robertsonian jumping translocation at amniocentesis: 45,XY,der(15;22)(q10;q10)mat/46,XY,i(15)(q10)/46,XY,genetic counseling,prenatal diagnosis and postnatal follow-up in a pregnancy with a favorable fetal outcome

ObjectiveWe present genetic counseling, prenatal diagnosis and postnatal follow-up of 45,XY,der(15;22)(q10;q10)mat/46,XY,i(15)(q10)/46,XY at amniocentesis in a pregnancy with a favorable fetal outcome.Case reportA 27-year-old, primigravid woman underwent amniocentesis at 19 weeks of gestation because increased nuchal translucency thickness, and the result was 45,XY,der(15;22)(q10;q10)[29]/46,XY,i(15)(q10)[3]/46,XY[5]. Simultaneous array comparative genomic hybridization (aCGH) analysis on the DNA extracted from uncultured amniocytes revealed arr (1–22) × 2, (X,Y) × 1. The maternal karyotype was 45,XX,der(15;22)(q10;q10), and the paternal karyotype was 46,XY. She was referred for genetic counseling, and repeat amniocentesis performed at 23 weeks of gestation revealed 45,XY,der(15;22)(q10;q10)mat[23]/45,XY,-22[2]. aCGH analysis on uncultured amniocytes detected no genomic imbalance, and polymorphic DNA marker analysis excluded uniparental disomy (UPD) 15. Fluorescence in situ hybridization (FISH) analysis using the chromosome 15q specific probe and the chromosome 22q specific probe detected three 15q signals in 4/104 cells (3.8%). The woman was advised to continue the pregnancy, and, a 3186-g phenotypically normal male baby was delivered at 38 weeks of gestation. The umbilical cord had a karyotype of 45,XY,der(15;22)(q10;q10) (40/40 cells). When follow-up at age seven months, the neonate was normal in development, the peripheral blood had a karyotype of 45,XY,der(15;22)(q10;q10) (40/40 cells), and the buccal mucosal cells had normal signals in all 100 cells.ConclusionsMosaicism for Robertsonian jumping translocations at amniocentesis can be a transient condition and can be associated with a familial Robertsonian translocation and a favorable fetal outcome. Prenatal diagnosis of a Robertsonian jumping translocation involving chromosome 15 should include UPD 15 testing to exclude UPD 15.     

Microphthalmia with linear skin defects (MLS) syndrome evaluated by prenatal karyotyping, FISH and array comparative genomic hybridization

OBJECTIVE: To explore the utility of comparative genomic hybridization to BAC arrays (array CGH) for prenatal diagnosis of microphthalmia and linear skin defects syndrome. METHODS: We used karyotype analysis, FISH and array CGH to investigate an X;Y translocation. Replication studies were done on cultured amniocytes and lymphoblasts. RESULTS: We describe a severe case of MLS syndrome that presented prenatally with multiple anomalies including cystic hygroma, microphthalmia, intrauterine growth restriction and a complex congenital heart defect. Cytogenetic analysis of amniocytes revealed an unbalanced de novo translocation between chromosomes X and Y [karyotype 46,X,der(X)t(X;Y)(p22.3;q11.2).ish der(X)(DXZ1+,DMD+,KAL-,STS-,SRY-),22q11.2 (Tuple1 x 2)]. MLS diagnosis was made at birth and the prenatal karyotype was confirmed. Replication studies showed the derivative X chromosome was the inactive X. Array CGH confirmed the X and Y imbalances seen in the karyotype and also showed twelve BACs in the MLS region were deleted as a result of the translocation. FISH with BAC clones verified the array findings and placed the X breakpoint in Xp22.2, resulting in the amended karyotype, 46,X,der(X)t(X;Y)(p22.2;q11.2).ish der(X)(DXZ1+,DMD+,KAL-,STS-,SRY-),22q11.2(Tuple1 x 2) arr cgh Xp22.33p22.2(LLNOYCO3M15D10 -->GS1-590J6)x 1,Yq11.222q23(RP11-20H21-->RP11-79J10)x 1. CONCLUSION: The sensitivity of array CGH was valuable in detecting monosomy of the MLS critical region. Array CGH should be considered for the prenatal diagnosis of this syndrome.     

Detection of mosaicism for 46,X,i(Y) (q10) in the blood lymphocytes in a phenotypically normal male neonate with prenatally detected 45,X/46, XY at amniocentesis and cytogenetic discrepancy in various tissues

ObjectiveWe present detection of mosaicism for 46,X,i(Y) (q10) in the blood lymphocytes in a phenotypically normal male neonate with prenatally detected 45,X/46, XY at amniocentesis and cytogenetic discrepancy in various tissues.Case reportA 35-year-old, gravida 2, para 1, woman underwent amniocentesis at 17 weeks of gestation because of advanced maternal age. Amniocentesis revealed a karyotype of 45,X [8]/46,XY [15]. Simultaneous array comparative genomic hybridization (aCGH) on uncultured amniocytes revealed the result of arr (Y) × 0–1 with 25.493-Mb mosaic deletion of chromosome Yp11.31-q11.23. Prenatal ultrasound findings were unremarkable. The fetus had normal male external genitalia on fetal ultrasound. Following genetic counseling, the pregnancy was carried to 38 weeks of gestation, and a phenotypically normal male baby was delivered without any abnormalities of the male external genitalia. The cord blood had a karyotypes of 46,X,i(Y) (q10)[8]/45,X[3]/46,XY [29], and placenta had a karyotypes of 45,X [25]/46,X,i(Y) (q10)[7]/46,XY [8]. When follow-up at age two months, the neonate was normal in development. The peripheral blood had a karyotypes of 46,X,i(Y) (q10)[8]/45,X[5]/46,XY [27]. Interphase fluorescence in situ hybridization (FISH) analysis on 101 buccal mucosal cells showed normal X and Y signals in 101/101 cells.ConclusionFetuses with 45,X/46, XY at amniocentesis can be associated with mosaicism for 46,X,i(Y) (q10) in the blood lymphocytes, cytogenetic discrepancy in various tissues and a favorable outcome.     

Spontaneous menstruation in patients with Turner syndrome in our observations

OBJECTIVES: Patients with Turner syndrome (TS) may present a wide spectrum of gonadal function including spontaneous menstruation and fertility. DESIGN: The aim of our study was to present the patients with Turner syndrome (TS) with spontaneous menstruations considering specific karyotype and X-inactivation processes. MATERIALS AND METHODS: 5 women from group of 55 patients in age from 15 to 38 years with diagnosis of TS and gonadal function were found. Clinical analysis included the evaluation of spontaneous pubertal development and hormones levels. Cytogenetic analysis was performed on peripheral blood samples using GTG banding technique. X inactivation studies were done by dynamic RBG technique. RESULTS: In two patients with mosaic karyotype and predominant 46,XX line two pregnancies were observed. They had regular menses and normal sexual development. In one patient (karyotype: 45,X[2]/46,XX[98]) spontaneous abortions and premature birth were present. Second patient was (46,XX[245]/46,X,r(X)(p22q26)[5]) in pregnancy in this time. Another three patients menstruated irregularly. The menarche appeared later. The karyotypes were: 46,X,del(X)(p11.3) in two patients and 45,X[64]46,X,r(X) (p22q26)[18]/46,XX[4] in one. CONCLUSIONS: We conclude that spontaneous menstruations and possibility of pregnancy depend on specific karyotype in patients with TS.