双色FISH和DNA纤维FISH方法的建立与应用

在构建了含毛细胞白血病相关的结构性倒位inv (5) (p13.1q13.3)的细胞系后,为了确定该新建细胞系在建株过程中其倒位断裂点关键区遗传物质是否发生改变,以生物素或地高辛标记的cCI5-216 和cCI5-267黏粒DNA为探针,进行染色体中期、间期和DNA纤维3种双色荧光原位杂交的分析。结果表明：该新建细胞系的3种双色荧光原位杂交结果,均与该细胞系的原代细胞的完全相同,证实了该细胞系倒位断裂点关键区的遗传物质结构未发生改变。该细胞系是揭示毛细胞白血病发病的分子机理的重要研究材料。     

染色体末端微小结构异常的分子细胞遗传检测

为检出易于被忽略的染色体末端微小结构异常,为生育提供指导,选取特异性7号全染色体探针,X染色体长臂探针和7q亚端粒（7q36→qter)探针,用荧光原位杂交（fluorescence in situ hybridzation,FISH)结合G显带技术分析2个病例,其中病例1有不良妊娠史并疑有末端微小易位,病例2在G显带水平已发现为X和7号染色体易位的卵巢早衰患者,结果表明,FISH确诊病例1为染色体末端的隐匿易位,病例2的易位断点得到精确定位,它不在7q36而在7q末端。应用特异性染色体探针及亚端粒探针,通过FISH技术可以确诊染色体末端区域的微小结构异常,在临床遗传学中有广泛的应用,是遗传咨询和生育指导的有效工具之一。     

335例遗传咨询病理的染色体分析

本文对335例遗传咨询病理的外周血淋巴细胞进行培养,并用G带、C带、Ag-NOR和高分辨G显带及基因探针进行原位杂交等技术进行了染色体分析。检出异常核型35例,占10.4%,其中3例较少见,2例国内未见报道。     

335例遗传咨询病理的染色体分析

本文对335例遗传咨询病理的外周血淋巴细胞进行培养，并用G带、C带、Ag-NOR和高分辨G显带及基因探针进行原位杂交等技术进行了染色体分析。检出异常核型35例，占10.4%，其中3例较少见，2例国内未见报道。     

4 号染色体部分重复和缺失一例

本文报道一例4号染色体部分重复和缺失的男婴。患儿发育迟缓,智力低下,体查多种畸形。其G显带核型为46,XY,rec(4),dupq,inv(4)(p16q31)。患儿父亲及祖母均为inv(4)(p16q31)的携带者,重复一缺失染色体源自患儿父亲的减数分裂重组。     

肿瘤染色体畸变分析方法新进展

摘要: 肿瘤的发生多与染色体畸变有关, 确定染色体畸变与肿瘤的关系, 必然离不开染色体畸变的检测分析。文章简要综述几种常用染色体畸变的检测方法及其新进展, 包括G显带、荧光原位杂交(FISH )、光谱核型分析(SKY)、多色荧光原位杂交(M-FISH)、多色显带分析技术(Rx-FISH)、比较基因组杂交(CGH)和微阵列比较基因组杂交(Array CGH), 以及这些方法在肿瘤诊断和研究方面的应用。     

多色-FISH技术的进展及其在分子生物医学上的应用

传统显带分析技术以每条染色体独特的显带带型为依据,提供染色体形态结构的基本信息,用于染色体核型的初步分析。然而有些染色体重排由于涉及的片断太小或具有相似的带型,用该方法难以探测或准确描绘。多元荧光原位杂交(M-FISH),光谱核型分析(SKY),FISH-显带分析技术是染色体特异的多色荧光原位杂交技术(mFISH)。它们能够探测出传统显带分析不能发现的染色体异常,提供更准确的核型。M-FISH和SKY均以组合标记的染色体涂染探针共杂交为基础,二者的不同在于观察仪器和分析方法上。它们可对中期染色体涂片进行快速准确分析,描绘复杂核型,确认标记染色体,主要用于恶性疾病的细胞遗传学诊断分析。FISH-显带分析技术以FISH技术为基础,能同时检测多条比染色体臂短的染色体亚区域。符合该定义的FISH-显带分析技术各有特点,其共同特点是都能产生DNA特异的染色体条带。这些条带有更多色彩,能提供更多信息。FISH-显带分析技术已经成功地被用于进化生物学、放射生物学以及核结构的研究,同时也被用于产前、产后以及肿瘤细胞遗传学诊断,是很有潜力的工具。     

白菜rDNA及C-(0)t-1DNA的荧光原位杂交及其核型分析

以早熟白菜苔为实验材料,从其基因组DNA中分离出C0t-1 DNA并用生物素标记作探针,25S rDNA用地高辛标记作探针,对有丝分裂中期相染色体进行双色荧光原位杂交。每对染色体上均显示出了特定的C0t-1 DNA荧光原位杂交带型,5对染色体上显示出了25S rDNA荧光原位杂交带型。双色荧光原位杂交证实了C0t-1 DNA与25S rDNA二者具有一致的染色体位置特征,表明基于rDNA及C0t-1 DNA的荧光原位杂交核型分析技术,优于目前普遍采用的只基于rDNA的荧光原位杂交核型分析方法。结合C0t-1 DNA与25S rDNA的荧光原位杂交带型和传统的染色体的形态学标记分析方法及白菜已公布的基于rDNA分布的核型分析结果,创建了一个精确的白菜核型。     

白菜rDNA及C0t-1DNA的荧光原位杂交及其核型分析

以早熟白菜苔为实验材料,从其基因组DNA中分离出C0t-1DNA并用生物素标记作探针,25SrDNA用地高辛标记作探针,对有丝分裂中期相染色体进行双色荧光原位杂交。每对染色体上均显示出了特定的C0t-1DNA荧光原位杂交带型,5对染色体上显示出了25SrDNA荧光原位杂交带型。双色荧光原位杂交证实了C0t-1DNA与25SrDNA二者具有一致的染色体位置特征,表明基于rDNA及C0t-1 DNA的荧光原位杂交核型分析技术,优于目前普遍采用的只基于rDNA的荧光原位杂交核型分析方法。结合C0t-1 DNA与25SrDNA的荧光原位杂交带型和传统的染色体的形态学标记分析方法及白菜已公布的基于rDNA分布的核型分析结果,创建了一个精确的白菜核型。     

荧光原位杂交技术检测骨髓增生异常综合征染色体异常的临床意义

目的:探讨荧光原位杂交(FISH)技术检测骨髓增生异常综合征(MDS)染色体异常的敏感性,特异性及临床意义。方法:采用细胞遗传学分析(CCA)和组合探针CSF1R/D5S23,D5S721(5q33),EGR1/D5S23,D5S721(5q31),D7S486/CSP7(7q31),D7S522/CSP7(7q31),D20S108/CSP8(20q12/CSP8)检测45例MDS患者骨髓细胞的染色体异常,并比较检测结果。结果:两种方法共检出染色体异常26例(58%),染色体数目异常9例,占34.6%;染色体结构异常13例,占50%;复杂核型4例。CCA检出+8和20q-各3例,7q-2例;FISH检出7号染色体异常8例占17.8%(8/45),两组间比较差异有统计学意义(P=0.0441713)。FISH检出+8和20q-各5例,5q-异常4例。7号染色体异常和复杂核型组与核型正常组比较转白率高。结论:组合探针检出MDS中5q-,-7/7q-,+8,20q-核型异常高于CCA,CCA结合FISH技术能提高MDS染色体异常的检出率,对于疾病诊断,判断预后具有重要价值。     

A prevalent Y;15 translocation in the Ethiopian Beta Israel community in Israel

We describe 7 cases of abnormal karyotypes involving chromosomes Y and 15 in Ethiopian Beta Israel patients: 46,XX, der(15)t(Y;15)(q12;p12) and 46,XY,der(15)t(Y;15)(q12;p12). Six cases were incidentally found in amniocentesis performed for various indications; the indication for karyotyping in 1 case was recurrent abortions. To the best of our knowledge, this is the first report of this translocation in a specific ethnic group. We conclude that the derivative chromosome 15 with chromosome Y is probably a normal variant in Ethiopian Beta Israel occurring at an estimated frequency of 4/74 (5.4%). The prenatal diagnosis of this translocation in this population probably does not require further parental testing.     

Paracentric inversion of Yq and review of the literature

We report on the second prenatal diagnosis of familial paracentric inversion of the long arm of Y chromosome [46, X, inv(Y)(q11.2q12)]. The anomaly was detected through an amniocentesis performed because of advanced maternal age. The inversion has been detected by standard GTG banding methods and better characterized by FISH with painting probe and specific satellite probes DYZ1 and DYZ3. The inversion derived from phenotypically normal father. Pregnancy was uneventful and an healthy child was born. We discuss the issue concerning genetic prenatal counselling of this rare condition and we report the clinical follow up of the child.     

A new type of inversion of a human Y chromosome.

Using chromosome banding techniques, a phenotypically normal male was found to have an abnormal banding pattern of the Y chromosome. By the constitutive heterochromatin staining method, a darkly stained band was located on the short arm and the proximal region of the long arm. The quinacrine staining method also showed a similar abnormal banding pattern: a brightly fluorescing band was seen on the short arm and the proximal region of the long arm. By the conventional Giemsa staining method, however, no specific morphological abnormality was detected in the aberrant Y. On detailed karyotype analyses no recognizable abnormality of banding patterns of any other chromosome was found aside from the abnormal Y. The abnormality was determined to be a complex inversion of the Y chromosome, which is described as 46,X,inv(Y)(pter leads to p11::q11 leads to q12::cen::q12 leads to qter).     

Deletion of specific sequences or modification of centromeric chromatin are responsible for Y chromosome centromere inactivation

Summary Stable dicentric chromosomes behave as monocentrics because one of the centromeres is inactive. The cause of centromere inactivation is unknown; changes in centromere chromatin conformation and loss of centromeric DNA elements have been proposed as possible mechanisms. We studied the phenomenon of inactivation in two Y centromeres, having as a control genetically identical active Y centromeres. The two cases have the following karyotypes: 45,X/46,X,i(Y)(q12) and 46,XY/ 47,XY,+t(X;Y)(p22.3;p11.3). The analysis of the behaviour of the active and inactive Y chromosome centromeres after Da-Dapi staining, CREST immunofluorescence, and in situ hybridization with centromeric probes leads us to conclude that, in the case of the isochromosome, a true deletion of centromeric chromatin is responsible for its stability, whereas in the second case, stability of the dicentric (X;Y) is the result of centromere chromatin modification.     

无精和严重少精症患者的遗传缺陷分析——-附2例世界首报染色体异常核型

对217例无精和严重少精症患者外周血淋巴细胞染色体核型进行分析,并采用聚合酶链反应对7例Y染色体结构异常患者的AZFc区进行检测。发现187例无精症患者中检出异常核型77例（41.18%）（其中46,XY,t(6;14)(p21;p13),46,XY,t(8;12)(p21;q24)为世界首报核型）,主要涉及染色体异常（数目异常和结构异常）;染色体异态（Y染色体异态和9号染色体臂间倒位）及46,XX性反转;30例严重少精症患者中检出异常核型4例（13.33%）（结构异常和46,XX性反转）。由此可见,性染色体数目和结构异常是精子发生障碍的主要原因,其次常染色体的某些断裂点也可能影响精子发生。AZFc区的缺失与否与精子发生也有直接关系。     

Acquired inversions in human leucocytes

Peri- and paracentric inversions are observed in human leukocytes at various rates. Four categories are proposed, in relation to the frequency of occurrence, although it may vary with time for a same inversion. Category 1 corresponds to isolated, thus non recurrent inversions. Category 2 (f congruent to .001) corresponds to inv(14)(q12qter) and inv(7)(p14q35) in individuals with presumably normal genetic constitution. Category 3 (f congruent to .01) corresponds to inv(7)(p14q35) in patients affected by ataxia telangiectasia (AT). This inversion, when it is frequent, indicates an abnormal genetic constitution, radiation sensitive and predisposing to cancers. Finally, category 4 (f greater than or equal to .1) corresponds to inversions existing in precancer or in cancer clonal cells: inv(14)(q11.2q32.2) in AT patients affected by a T-cell hemopathy, inv(14)(q12qter) in chronic T-cell lymphocytic leukaemia, and inv(16)(p13q22) in acute myelomonocytic leukaemia with abnormal eosinophils. The prognostic and diagnostic interests of these inversions is discussed.     

罗伯逊易位分子机理初探

目的：探讨复发性自然流产与染色体罗伯逊易位间的关系。方法：采用人外周血淋巴细胞培养,常规G显带技术行染色体核型检查,并结合临床资料对其进行分析。结果：57对复发性自然流产夫妇中,检出罗伯逊易位染色体核型4例,检出率3.51%。结论：罗伯逊易位是导致复发性流产的重要原因之一,对复发性自然流产患者进行常规的染色体检查及遗传咨询具有一定的临床意义。     

Cytogenetic evaluation of 163 azoospermics

A constitutional chromosomal aberration was diagnosed in 38/163 (23.3%) azoospermic patients. Whereas the 47,XXY complement was the commonest (31/38 cases), the following abnormal karyotypes were also found: 46,XX; 46,X,del(Y) (q11); 46,X,r(Y); 46,XY,inv(1) (p3500q21.3)mat; and 46,Y,t(X;3) (q26;q13.2)mat (both the deleted and the annular Y were observed twice). Pooled data from the literature showed that the frequency of chromosomal abnormalities is higher in azoospermic (150.4/1000) than in infertile (55.3/1000) males, which in turn is higher than in newborns (less than 6/1000). The observed different frequency between azoospermic and infertile individuals is given by several types of chromosomal abnormalities, mainly by the complement 47,XXY. The analysis also showed that the male infertility secondary to rob translocations and supernumerary marker chromosomes is usually not related to azoospermia. The contrary occurs in certain rcp and gonosome;autosome translocations and in autosome inversions.     

Heterogeneity of pericentric inversions of the human y chromosome

Pericentric inversions of the human Y chromosome (inv(Y)) are the result of breakpoints in Yp and Yq. Whether these breakpoints occur recurrently on specific hotspots or appear at different locations along the repeat structure of the human Y chromosome is an open question. Employing FISH for a better definition and refinement of the inversion breakpoints in 9 cases of inv(Y) chromosomes, with seemingly unvarying metacentric appearance after banding analysis, unequivocally resulted in heterogeneity of the pericentric inversions of the human Y chromosome. While in all 9 inv(Y) cases the inversion breakpoints in the short arm fall in a gene-poor region of X-transposed sequences proximal to PAR1 and SRY in Yp11.2, there are clearly 3 different inversion breakpoints in the long arm. Inv(Y)-types I and II are familial cases showing inversion breakpoints that map in Yq11.23 or in Yq11.223, outside the ampliconic fertility gene cluster of DAZ and CDY in AZFc. Inv(Y)-type III shows an inversion breakpoint in Yq11.223 that splits the DAZ and CDY fertility gene-cluster in AZFc. This inversion type is representative of both familial cases and cases with spermatogenetic impairment. In a further familial case of inv(Y), with almost acrocentric morphology, the breakpoints are within the TSPY and RBMY repeat in Yp and within the heterochromatin in Yq. Therefore, the presence of specific inversion breakpoints leading to impaired fertility in certain inv(Y) cases remains an open question.     

Familial pericentric inversion of the X chromosome [inv(X)(p11q28)]

A familial pericentric inversion of the X chromosome [46,X,inv(X)(p11q28)] and [46,inv(X)(p11q28), Y] is reported. The carriers of the inv(X) presented no clinical symptoms. Either the inverted or the normal X chromosome may be late replicating.