自然流产夫妇的细胞遗传学研究─附10例世界首报染色体核型

用人体外周血淋巴细胞培养法对６２４对原因不明的自然流产夫妇进行了细胞遗传学研究，检出异常染色体携带者５７例，其中女３８例，男１９例。５７例染色体异常中，结构异常４５例；染色体数目异常２例；９号染色体臂间倒位９例，Ｙ染色体倒位１例。异常染色体涉及到１，３，４，５，６，７，８，９，１０，１１，１３，１４，１５，１７，１８，１９，２１，２２，Ｘ，Ｙ共２０条。４５例结构异常中１０例经湖南医科大学国家细胞遗传学培训中心鉴定为世界首报。     

2269例细胞遗传学研究

总结Ｔ２２６９例的细胞遗传学资料，发现染色体异常３１０例，总异常率为１３，６６％。各组中智力低下的染色体异常率最高，占４４．４４％，大多为２１三体；男、女生殖缺陷的染色体异常率次之，分别为２７．７８％和３５．８１％，主要为性染色体异常；流产及异常生育史的染色体异常率最低，分别为４，１４％和１．１０％，主要为常染色体平衡易位。提示细胞遗传学检查对上述疾病的早期诊断和早期治疗有重要意义。     

荧光原位杂交技术在细胞遗传学中的应用

目的：探讨荧光原位杂交技术（ＦＩＳＨ）在细胞遗传学异常染色休核型分析中的应用价值。方法：应用Ｘ，Ｙ，１３／２１，１８号染色体α卫星ＤＮＡ探针（包括生物素标记及地高辛标记的探针，）对原Ｇ带核型为４６，ＸＯ＋ｒ？）；４６，ＸＯ＋ｉ（Ｘｐ）９７％／４７，ＸＯ＋２ｉ（Ｘｐ）３％；４７．ＸＸＸ／４５，ＸＯ；４７，ＸＹ＋２１等４例的外周血染色体及间期细胞进行原位杂交，并用正常男、女核型作为阳性对照，以不加探针的杂液反应为阴性对照，杂交后用ＯｌｙｍｐｕｓＢＸ６０荧光显微镜观察玻片并照像。结果：原Ｇ带核型４７，ＸＹ＋２１；４７，ＸＸＸ／４５，ＸＯ与ＦＩＳＨ结果一致，分别诊断为先天性愚型及Ｔｕｒｎｅｒ综合征。原Ｇ带核型４６，ＸＯ＋ｒ？的ｒ是征，其真正核型是４６，ＸＯ＋ｄｉｃ（Ｙｑ）／４７，ＸＯ＋２［ｄｉｃ，ｉ（Ｙｑ）］。正确诊断为Ｙ染色体结构异常与性腺发育不全综合征。结论：对一些常规Ｇ带难以确诊的复杂染色体结构畸变核型。如环状、易位等，ＦＩＳＨ技术具有非常重要的效用。     

性畸形四例遗传学分析

在细胞遗传学核型分析的基础上，利用Ｙ染色性体决定区基因（ｓｅｘ－ｄｅｔｅｒｍｉｎｉｇｒｅｇｉｏｎｏｆｔｈｅＹｃｈｒｏｍｏｓｏｍｅ，ＳＲＹ）探针，对４例性畸形病例进行了Ｓｏｕｔｈｅｒｎ杂交分析。同时运用ＳＲＹ编码区的特异性引物，对其基因组ＤＮＡ进行了聚合酶链式反应（ＰＣＲ）扩增检测。结果显示：１例４６，ＸＹ女性伴性腺发育不良症；１例４５，ＸＯ／４６，ＸＹ女性表现为Ｔｕｒｎｅｒ综合征，未检出ＳＲＹ特异     

502对反复自然流产夫妇的染色体研究

对反复自然流产２次或２次以上的５０２对夫妇进行外周血培养Ｇ显带染色体核型分析。发现异常核型５２例，占５．１８％；其中平衡易位３８例，占７３．０８％；染色体数目异常５例，占９．６２％；嵌合体８例，占１５．３８％；９号染色体臂间倒位１例，占１．９２％。结果表明，平衡易位携带者是反复自然流产夫妇中最常见的染色体异常，故流产史是检出人群中平衡易位携带者的重要临床指征。     

性分化异常患者的细胞遗传学病因分析

目的：性分化异常的细胞遗传学病因。方法：每例性分化异常患者取外周血淋巴细胞常规制备染色体。Ｇ带、Ｃ带,高分辨显带及银染技术分析核型。结果：１１１例性分化异常患者中检出染色体异常４５例,数量异常１４例,结构异常３１例,包括缺失、倒位、易位、环状染色体、额外小染色体等。结论：染色体异常是性分化异常的重要原因,分析患者的核型,对于诊断及治疗均有一定意义。     

性分化异常患者的细胞遗传学分析

目的：探讨性分化异常的遗传学病因。方法：每例性分化异常患者取外周血淋巴细胞常规制备染色体。Ｇ带、Ｃ带，高分辨显带及银染技术分析核型。结果：１１１例性分化异常患者中检出染色体异常４５例，数量异常１４例，结构异常３１例，包括缺失、倒位、易位、环状染色体、额外小染色体等。结论：染色体异常是性分化异常的重要原因，分析患者的核型，对于诊断及治疗均有一定意义。     

妊娠发育停止妇女绒毛膜细胞遗传学特征

为研究能导致胎卵发育停止的染色体异常数量及结构，比较临床上普遍应用的两种细胞遗传学检查方法所显示染色体异常的信息量，对630例早期妊娠胎卵发育停止妇女子宫刮出物中的叶状绒毛膜绒毛标本进行前瞻性细胞遗传学研究。630个绒毛膜标本中可供染色体分析的582个（除外48个无明确细胞遗传学结论的标本）。     

经母血采集胎儿细胞行产前诊断的最佳时间探讨

目的：探讨利用母血循环中胎儿细胞进行产前诊断的最佳采血时间。方法：对４１例孕龄为６～１４周的妇女连续取血，采用套式聚合酶链反应技术检测人类Ｙ染色体特异的锌指蛋白基因（ＺＦＹ）。结果：１９例妊娠男性胎儿妇女外周血ＺＦＹ随着孕龄的增加，其胎儿单拷贝基因的检出率增高，其中孕６周时检出率为１／１９（５．３％），孕１１周时为１３／１９（６８．４％），而到孕１４周时，则达到１８／１９（９５．０％）；对２２例妊娠女性胎儿妇女外周血进行ＺＦＹ检测时，无一例假阳性结果，这一检测方法在妊娠早期进行胎儿性别鉴定的总准确率达到９７．８％（４０／４１）。结论：利用母血循环中胎儿细胞进行产前诊断的最佳采血时间应在妊娠１４周，同时提示胎儿细胞最早进入母血循环中的时间在不同个体间存在明显的差异。     

反复流产夫妇的细胞遗传学研究

反复流产夫妇的细胞遗传学研究善志红，沈汝端，张兰，高桦细胞遗传学研究发现，染色体异常是反复流产发生的重要原因之一。本研究对３９０对反复流产夫妇的染色体进行检查，分析染色体异常与流产次数的相关性。一、资料与方法１．资料：自１９８３年至１９９３年，通过遗...     

Cytogenetic investigations in couples with repeated miscarriages and malformed children: report of a novel insertion

Recurrent miscarriage and the birth of a malformed child continue to be challenging problems for the patient and clinician. Often, cytogenetic studies have an important role in the evaluation of couples with a poor obstetric history. The present study deals with analysis of chromosome abnormalities in 2150 couples from India, and is the largest group ever reported in the literature. The observed incidence of chromosomal rearrangements in these couples was 3.5%. Apart from reciprocal, Robertsonian, inversions, a unique case of chromosome insertion was identified, which is perhaps only the second report in literature. Minor chromosome variants such as inv(9),inv(Y),9qh+, D/G variants were observed in 108 subjects. Cytogenetic studies should be performed for all couples with repeated miscarriages and bad obstetric history, and in cases of detected chromosomal aberration, the patient should be counselled individually according to the type of anomaly.     

Role of FISH on uncultured amniocytes for the diagnosis of aneuploidies in the presence of fetal anomalies

OBJECTIVE: To assess the accuracy of fluorescent in situ hybridization (FISH) on amniocytes in fetuses affected by structural malformations suggestive of chromosomal anomalies. METHODS: FISH of uncultured amniotic fluid cells and conventional cytogenetic analysis were performed on 48 pregnancies with ultrasonographic (US) evidence of fetal anomalies. The AneuVysion assay (Vysis) with specific probes for chromosomes 13, 18, 21, X and Y, was used. Amniotic fluid samples were obtained between the 14th and 34th weeks of gestation. RESULTS: In cases with a single abnormal US finding (n = 15), 5 aneuploidies were detected (1 case of trisomy 13 and 4 of trisomy 21). In the group with two or more malformations (n = 33) there were 15 aneuploidies (9 cases of trisomy 18, 2 of trisomy 21, 2 monosomy X, 1 trisomy 13, and 1 triploidy). In this group, conventional cytogenetic analysis revealed two additional chromosomal anomalies not detectable by FISH (1 trisomy 16 mosaic, and a terminal deletion 4p). No sex aneuploidies were observed. CONCLUSIONS: The lack of false-positive diagnosis in the FISH analysis in our sample prompts us to consider interphase FISH as a useful tool in pregnancies at high risk for chromosomal aneuploidies. When FISH analysis is normal, the overall risk of chromosomal abnormalities is significantly reduced. However, the finding of two chromosomal anomalies undetectable by AneuVysion assay confirms the need for conventional chromosome analysis to complement FISH results. Moreover, the results collected here, in agreement with those already reported in the literature, indicate that FISH analysis on uncultured amniocytes can play an important role in counselling and decision-making, especially in cases at risk for aneuploidies, such as those with structural abnormalities at US.     

576例先天畸形儿的染色体核型分析

目的：通过对超声确诊为先天畸形儿的染色体核型进行分析,总结染色体异常与畸形部位和畸形数目的关系,为产前诊断提供依据。方法：2009年1月—2013年12月在广西壮族自治区妇幼保健院行羊水穿刺或脐带血穿刺,产前诊断指征为先天畸形儿的576例胎儿进行染色体核型分析,探讨先天畸形儿与染色体异常的关系及比例。结果：细胞培养成功551例（成功率95.66%）。染色体异常90例（16.33%）,其中常染色体异常63例（占染色体异常的70.00%）,性染色体异常25例（占染色体异常的27.78%）,三倍体1例,标记染色体1例。 染色体异常的畸形儿中,主要为心脏畸形31例（34.44%）,皮肤水肿30例（33.33%）,面颈部异常21例（23.33%）,脑部结构异常20例（22.22%）,消化系统畸形11例（12.22%）。结论：先天畸形儿的染色体异常发生率极高;对超声提示为先天性畸形儿者应行介入性产前诊断,以避免染色体异常患儿的出生。     

Cytogenetic studies in patients with reproductive failure

BACKGROUND: Cytogenetic studies in patients with reproductive failure AIM: To investigate the contribution of chromosomal abnormalities in sub fertility and in couples with repeated abortions. METHODS: Hundred and 13 couples who had at least two or more spontaneous abortions and 65 women and 63 men with infertility were analyzed cytogenetically. RESULTS: Major chromosomal rearrangements were found in 8% and minor variants in 6% in the study population. Major chromosomal aberrations were judged to explain 4.9% of recurrent abortions and 13% of infertility. Chromosomal abnormalities in infertile men occurred in 5% and in infertile women in 21.5%. The chromosomal abnormalities were structural (57%), numerical (18%) or mosaics (25%). CONCLUSIONS: Chromosomal aberrations in recurrent abortions are mostly structural ones and those in female infertility mosaicism of sex chromosomes. Turner's syndrome, Turner variants and XY females are detected as a cause of female infertility. The structural and numerical aberrations of either sex or autosomal chromosomes were found in infertile men.     

Cytogenetic analysis of 179 Iranian women with premature ovarian failure

Abstract

The importance of chromosomal abnormalities in etiology of premature ovarian failure (POF) is well known but in many cases, POF still remains idiopathic. We investigated the frequency and type of chromosomal aberrations in Iranian women diagnosed with idiopathic POF. Standard cytogenetic analysis was carried out in a total of 179 patients. Karyotype analysis of these patients revealed that 161 (89.95%) patients had normal female karyotype and 18 (10.05%) patients had abnormal karyotypes. The abnormal karyotypes included sex reverse sex determining region Y (SRY) negative (five Cases), X chromosome mosaicism (five cases), abnormal X chromosomes (three cases), abnormal autosomes (three cases) and X-autosome translocation (two cases). The overall prevalence of chromosomal abnormalities was 10.05% in this first large-scale report of chromosomal aberrations in Iranian women with POF. The results confirm previous observations and emphasis on the critical role of X chromosome abnormalities as one of the possible etiologies for POF.     

Effectiveness of prenatal chromosomal analysis using multicolor fluorescent in situ hybridisation

Objective To evaluate the clinical effectiveness of multicolour fluorescent in situ hybridisation (FISH) analysis in routine prenatal diagnosis.
Design Prospective study.
Sample 3203 amniotic fluid samples.
Methods Unique DNA (chromosomes 13 and 21) and α satellite centromeric-specific (chromosomes X, Y and 18) probes were used in two mixes to permit the simultaneous analysis of several chromosomes. The performance of multicolour FISH and conventional cytogenetic analysis was compared.
Results Conventional cytogenetic analysis identified 111 chromosomal abnormalities, of which 94 were potentially detectable by the FISH technique and 97 would be typically associated with neonatal phenotypic abnormalities. Multicolour FISH analysis detected 84% (93/111) of all chromosome abnormalities and 99% (93/94) of abnormalities where there was a specific probe. The sensitivity of multicolour FISH analysis was 95% (92/97) for chromosomal abnormalities likely to result in an abnormal postnatal outcome. Multiple ultrasound abnormalities were detected in all five cases of clinically relevant chromosomal abnormalities missed by multicolour FISH. FISH results were available within 48 hours and the sample failure rate was 0.1% (3/3202).
Conclusion Multicolour FISH analysis is a sensitive and reliable technique for the rapid prenatal diagnosis of chromosomal abnormalities. Examining only five chromosomes allowed 95% of clinically relevant chromosomal abnormalities to be diagnosed correctly. As routine antenatal screening is targeted at the major autosomal trisomies and sex chromosome aneuploidies, multicolour FISH analysis may potentially replace conventional cytogenetic analysis in routine prenatal diagnosis.     

Rapid prenatal diagnosis of trisomy 21 in 5049 consecutive uncultured amniotic fluid samples by fluorescence in situ hybridisation (FISH).

OBJECTIVES: This was a retrospective study on the results of interphase fluorescence in situ hybridization (FISH), performed routinely for chromosome 21 and on ultrasonographic indications for chromosomes 13, 18, X and Y in a series of 5049 amniotic fluid samples. METHODS: Interphase FISH for chromosome 21 was performed in 5049 consecutive amniotic fluid samples for the rapid prenatal diagnosis of Down syndrome. Aneuploidy for four other chromosomes (13, 18, X and Y) was tested following ultrasonographic indications. Karyotypes from standard cytogenetic analysis were compared to the FISH results. RESULTS: Using conventional cytogenetics 3.6% (183/5049) chromosomal anomalies were detected. After exclusion of familial chromosome rearrangements, i.e. balanced autosomal reciprocal or Robertsonian translocations (30/5049) and inversions (19/5049), 2.65% chromosomal anomalies (134/5049) were diagnosed. Of this group 0.18% (9/5049) were chromosomal rearrangements not detectable by FISH and 2.47% (125/5049) were numerical chromosomal anomalies detectable by interphase FISH for chromosomes 13, 18, 21, X and Y. With routine interphase FISH for chromosome 21 and FISH on echographic indication for the other four chromosomes we detected 107/125 of these numerical chromosomal anomalies, i.e. 85.6%. All 70 cases of trisomy 21 were detected by FISH and confirmed with conventional cytogenetics (sensitivity=100%) and there were no false-positive results (specificity=100%). Maternal cell contamination of amniotic fluid samples occurred in 1.27% (64/5049) of samples; 0.26% (13/5049) of these samples were uninformative by FISH due to maternal cell contamination (12/5049) or absence of nuclei in one sample (1/5049). CONCLUSION: In this group of 5049 samples we found that FISH is a reliable technique for the rapid prenatal diagnosis of trisomy 21. The number of uninformative cases due to maternal cell contamination was low. The strategy to perform FISH for chromosome 21 in all samples and only on ultrasonographic indication for the four other chromosomes (13, 18, X and Y) followed by standard cytogenetics is effective.     

复发性流产夫妇的染色体结构异常分析

目的:探讨复发性流产与染色体结构异常的关系。方法:对有复发性自然流产史的112对夫妇进行外周血染色体核型分析。结果:112对夫妇中有20例染色体核型异常,异常检出率8.9％。其中平衡易位4例,复杂易位1例,臂间倒位8例,Y染色体变异6例,占2.7％;常染色体变异1例。结论:复发性流产夫妇任何一方的染色体结构异常均可引起流产等不良妊娠,9号染色体倒位和Y染色体变异与早期流产的关系密切。