Rapid detection of aneuploidy in uncultured chorionic villus cells using fluorescence <i>in situ</i> hybridization

Rao, P. Nagesh; Hayworth, R.; Cox, Kelly; Grass, Frank S.; Pettenati, Mark J.

doi:10.1002/pd.1970130402

Cited by 27 publications

(19 citation statements)

References 13 publications

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“…A striking example involves chromosomes 13 and 21, for which this sequence homology reaches 99.7% [Jorgensen et al, 1987;Charlieu et al, 1992]. Uncoupling between hybridized signal numbers and the number of target chromosomes was also reported [Lebo et al, 1992;Nagesh Rao et al, 1993].…”

Section: Introductionmentioning

confidence: 92%

Rapid chromosome detection by PRINS in human sperm

2001

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Primed in situ (PRINS) labeling may be used for direct estimation of disomy rate in human sperm. We combined the PRINS procedure with a rapid and efficient NaOH pretreatment allowing simultaneous decondensation and denaturation of sperm nuclei. This enabled double labeling of human sperm within a 2-hr time span. Chromosome-specific primers have been defined for most human chromosomes, including those most frequently involved in aneuploidy. Using the dual-color PRINS method, we estimated incidences of disomy for 15 autosomes and the sex chromosomes in sperm samples from 6 normal fertile men. The frequency of disomy ranged from 0.28% to 0.36%. There were no significant interchromosomal differences in disomy rate, but chromosome 21 displayed a higher incidence of nondisjunction than did the other chromosomes.

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Section: Introductionmentioning

confidence: 92%

Rapid chromosome detection by PRINS in human sperm

2001

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“…Fluorescence in situ hybridization (FISH) with chromosome‐specific probes can be applied for the detection of chromosome number aberrations in metaphase spreads, as well as in interphase nuclei [1–3]. FISH in the interphase nuclei is especially useful for clinical applications when rapid results are desirable and cells are difficult to culture in vitro to get analyzable mitoses [4].…”

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confidence: 99%

Optimization of the fluorescence in situ hybridization (FISH) technique for high detection efficiency of very small proportions of target interphase nuclei

et al. 2000

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Using commercially available fluorochrome-labeled probes specific for chromosomes X, Y, 13, 18, and 21, we optimized the technical protocols for fluorescence in situ hybridization (FISH) so that the highest sensitivity and specificity were achieved. Also, we compared the optical properties of different types of fluorescent labels in an effort to develop the most efficient FISH protocol, including the determination of which types of labels are the easiest to count accurately. The lymphocytes were purified from blood of normal male and female newborns, normal male and female adults, and a trisomy 21 male adult. Male and female lymphocytes were mixed in five different combinations. For each combination, the male lymphocytes either from newborns or from adults were diluted with female lymphocytes in seven different proportions. For each of these 35 different cell mixtures, 100,000 nuclei were analyzed and scored in a blind fashion. Among the different fluorochrome-labeled probes, the highest sensitivity and specificity were achieved when SpectrumAqua CEP-Y/SpectrumOrange CEP X probe mixture, SpectrumAqua CEP-18, SpectrumOrange LSI-13, and SpectrumOrange LSI-21 were hybridized. The hybridization sensitivity and specificity were higher than 99% for the identification of chromosomes X, Y, 13, and 18, and higher than 98% for the detection of trisomy 21. The proportion of false-positive signals was under 0.005% for XY detection and lower than 0.14% for autosome detection. With these high hybridization sensitivities and specificities, the optimized FISH protocol developed in our laboratory has the potential to detect very rare events, e.g., when the proportion of cells being sought is lower than 0.01%. In other words, our protocol allows the specific detection of one male cell sunken among 10,000 female cells.

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“…Also, uncoupling between hybridized signal numbers and the number of target chromosomes was sometimes reported (Lebo et al, 1992;Nagesh Rao et al, 1993).…”

Section: Introductionmentioning

confidence: 96%

PRINS as a method for rapid chromosomal labeling on human spermatozoa

Pellestor

Girardet

Lefort

et al. 1995

Molecular Reproduction Devel

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Direct in situ labeling of human spermatozoa was performed using the PRINS method. This technique is based on annealing of specific oligonucleotide primers, and subsequent primer extension by a Taq DNA polymerase. The reaction was carried out on a programmable temperature cycler, and labeling was obtained in a 1-hr reaction. The method was successfully tested with specific primers for chromosomes 13, 16, and 21. This suggests that PRINS may be a fast and reliable technique for detecting aneuploidies.

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Rapid detection of aneuploidy in uncultured chorionic villus cells using fluorescence in situ hybridization

Cited by 27 publications

References 13 publications

Rapid chromosome detection by PRINS in human sperm

Rapid chromosome detection by PRINS in human sperm

Optimization of the fluorescence in situ hybridization (FISH) technique for high detection efficiency of very small proportions of target interphase nuclei

PRINS as a method for rapid chromosomal labeling on human spermatozoa

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