Feasibility study of repeated fluorescent in-situ hybridization in the same human blastomeres for preimplantation genetic diagnosis

In order to increase the number of chromosomes examined in each blastomere, we have developed a repeated fluorescent in-situ hybridization (FISH) procedure by which six or more chromosomes can be analysed per blastomere of a human embryo. Three consecutive FISH procedures with directly-labelled fluo...

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Bibliographic Details
Published in:Molecular human reproduction 1998-10, Vol.4 (10), p.972-977
Main Authors: JIAEN LIU, TSAI, Y.-L, ZHENG, X.-Z, YAZIGI, R. A, BARAMKI, T. A, COMPTON, G, KATZ, E
Format: Article
Language:English
Subjects:
Adult
Biological and medical sciences
Biotechnology
Blastomeres - physiology
Cell Nucleus
Chromosomes, Human
Female
Fertilization in Vitro
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Humans
In Situ Hybridization, Fluorescence - methods
In vitro gene amplification. Pcr technique
Lymphocytes
Male
Methods. Procedures. Technologies
Oocytes - physiology
Pregnancy
Preimplantation Diagnosis - methods
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Summary:In order to increase the number of chromosomes examined in each blastomere, we have developed a repeated fluorescent in-situ hybridization (FISH) procedure by which six or more chromosomes can be analysed per blastomere of a human embryo. Three consecutive FISH procedures with directly-labelled fluorescent Vysis DNA probes were carried out for examination of chromosomes X, Y, 11, 13, 18 and 21 in the same blastomeres (n = 126) and lymphocytes (n = 164). Based on the initial number of nuclei, the percentages of nuclear loss and presence of signals were 3 and 92% respectively in blastomeres; 6 and 91% respectively in lymphocytes after the first FISH; 7 and 87% respectively in blastomeres and 10 and 86% respectively in lymphocytes, after the second FISH. These percentages were 13 and 78% respectively in blastomeres and 14 and 81% respectively in lymphocytes after the third FISH. The FISH procedure was repeated successfully in a couple for preimplantation genetic diagnosis of chromosomal aneuploidies in biopsied blastomeres of their embryos in our clinic. In conclusion, it is feasible to carry out repeated FISH procedures in the same blastomeres. Six or more chromosomes of a single blastomere may be examined using this procedure.
ISSN:1360-9947
1460-2407
1460-2407
DOI:10.1093/molehr/4.10.972