Analysis of chromosome 21 copy number in uncultured amniocytes by fluorescence <i>in situ</i> hybridization using a cosmid contig

Zheng, Yun‐Ling; Ferguson‐Smith, Malcolm A.; Warner, Jon; Erguson-Smith, M. E.; Sargent, Carole A.; Carter, N. P.

doi:10.1002/pd.1970121113

Cited by 44 publications

(41 citation statements)

References 18 publications

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“…Hybridization with the chromosome 21 contig probe, which is 55 kb in length and maps to 21q22.3 (Zheng et al 1992), produces clear signals in the form of spots, allowing the identification of the 21 bivalent (Figure 5a & b). The number of spots varies from 1 (with single spots usually being large) to 9 (with multiple spots usually being smaller) (see Table 1).…”

Section: Analysis Of Meiosis I Human Spermatocytesmentioning

confidence: 98%

Combined immunocytogenetic and molecular cytogenetic analysis of meiosis I human spermatocytes

Barlow

Hultén

1996

Chromosome Res

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We have used a combination of immunocytogenetic and molecular cytogenetic technology on human spermatocytes to investigate (1) meiosis I chromosome pairing, and (2) organization of synaptonemal complex (SC)-associated chromatin with respect to whole chromosome paints, unique DNA sequences and repetitive DNA of heterochromatic blocks, centromeres and telomeres. It is evident that synapsis normally starts at the termini of homologues. In general, synapsis proceeds synchronously from termini towards the centre of bivalents without any indication of interstitial initiation. Some aberrant meiosis I spermatocytes showed asynchronous pairing, demonstrating not only large differences in the degree of SC formation between bivalents, but also chromosome alignment without synapsis as well as clear interstitial synaptic initiation. It may be the case that alignment normally takes place along the entire length of homologues before synapsis occurs and that the potential for synaptic initiation exists along the length of chromosomes. Telomeric sequences were seen tightly associated with the SCs, as might be expected considering their kinetic properties in relation to the nuclear membrane. Other repetitive DNA, i.e. centromeric alpha-satellites and classical satellites of the heterochromatic blocks 1qh and 9qh, were all found to form loops that are associated with SCs only at their bases. A unique DNA cosmid probe (21q22.3) was found to produce a hybridization pattern consisting of spots located outside SC. The fluorescence in situ hybridization (FISH) signals of these spread DNA sequences have a granular appearance, probably reflecting the pattern of coiling and chromatin condensation of the target DNAs.

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Section: Analysis Of Meiosis I Human Spermatocytesmentioning

confidence: 98%

Combined immunocytogenetic and molecular cytogenetic analysis of meiosis I human spermatocytes

Barlow

Hultén

1996

Chromosome Res

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“…FISH was carried out in 26 cases, originally using a mixture of commercially available chromosome X-specific probe DXZ1 and the Y-specific heterochromatin probe DYZ1, directly labeled with fluorescein isothiocynate (FITC) (Oncor, Inc.), according to the procedure described by Zheng et al [6]. The mixture of X-and Y-specific probes and cell DNA were denatured simultaneously at 80°C for 10 min and hybridization was carried out overnight at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Improved Specificity of NRBC Detection in Chorionic Villus Sample Supernatant Fluids Using Anti-Zeta and Anti-Epsilon Monoclonal Antibodies

Mavrou

Kοlialexi²,

Zheng³

et al. 1999

Fetal Diagn Ther

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Objective: Fetal erythrocytes leak from fetal capillaries at the time of chorionic villus sampling (CVS). It has been reported that in approximately 60% of CVS cases fetal nucleated red blood cells (NRBC) can be isolated from the supernatant fluid by immunophenotyping with monoclonal antibody (Ab) against the γ-chain of fetal hemoglobin and used as an additional source for confirmation of the fetal karyotype. However, the increased prevalence of β-thalassemia mutations in countries such as Greece results in many pregnant women who produce γ-positive cells. This makes it difficult to distinguish between the fetal and maternal origin of the NRBC. Use of Abs against embryonic hemoglobin chains ζ and ε may increase specificity for fetal NRBC detection. Methods: Mouse monoclonal Abs against Hb-ζ and Hb-ε were used in order to examine if specificity for fetal NRBC detection in CVS supernatant fluids could be improved. 41 samples were studied using anti-ζ and 20 using anti-ε monoclonal Abs. Results: Anti-ζ or anti-ε positive erythrocytes were, respectively, identified in 52 of 61 CVS samples and anti-ζ or anti-ε positive NRBC were present in all cases. The mean number of Hb-positive erythrocytes identified with the anti-ζ Ab was 58 and the mean number of NRBC 29. The mean number of anti-ε positive erythrocytes was 30 and of NRBC 23. FISH with X and Y chromosome specific probes was performed in 26 cases and the results were concordant with the CVS karyotype. Statistical analysis using the correlation test showed that anti-ζ and anti-ε were more specific for the detection of embryonic NRBCs. Conclusions: Since embryonic monoclonal Abs show increased specificity, they should be preferentially used for NRBC detection in CVS supernatant fluids. Furthermore, the increased specificity of anti-ζ and anti-ε Abs may considerably improve prenatal diagnosis from fetal cells isolated from maternal circulation.

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“…Fluorescence in situ hybridization (FISH) to interphase nuclei with chromosome-specific DNA probes can now rapidly and accurately detect the most common autosomal trisomies and aneuploidies of the sex chromosomes (Cremer et al 1986;Lichter et al 1988aLichter et al , 1988bKlinger et al 1990Klinger et al , 1992 Christensen et al 1992; Lebo et al 1992;Zahed et al 1992;Zheng et al 1992). The general strategy for utilization of FISH has been extensively reviewed (McNeil et al 1991;Tkachuk et al 1991;Trask 1991;Ledbetter 1992;Sawyer et al 1992).…”

Section: Introductionmentioning

confidence: 99%

Rapid Prenatal Diagnosis of Chromosomal Aneuploidies by Fluorescence in Situ Hybridization: Clinical Experience With 4500 Specimens

Ward

Gersen

Carelli

et al. 1994

Obstetrical & Gynecological Survey

144

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Analysis of chromosome 21 copy number in uncultured amniocytes by fluorescence in situ hybridization using a cosmid contig

Cited by 44 publications

References 18 publications

Combined immunocytogenetic and molecular cytogenetic analysis of meiosis I human spermatocytes

Combined immunocytogenetic and molecular cytogenetic analysis of meiosis I human spermatocytes

Improved Specificity of NRBC Detection in Chorionic Villus Sample Supernatant Fluids Using Anti-Zeta and Anti-Epsilon Monoclonal Antibodies

Rapid Prenatal Diagnosis of Chromosomal Aneuploidies by Fluorescence in Situ Hybridization: Clinical Experience With 4500 Specimens

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