BRIEF REPORT |
Correspondence to: Jingly Fung, Dept. of Obstetrics, Gynecology and Reproductive Science, Univ. of CaliforniaSan Francisco, 533 Parnassus, San Francisco, CA 94143-0720.
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Summary |
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Chromosome abnormalities are common causes of congenital malformations and spontaneous abortions. They include structural abnormalities, polyploidy, trisomy, and mosaicism. In in vitro fertilization (IVF) programs, preimplantation genetic diagnosis (PGD) of oocytes and embryos has become the technique of choice to select against abnormal embryos before embryo transfer. For diagnosis of structural abnormalities, we developed case-specific breakpoint-spanning DNA probes. Screening of an in-house yeast artificial chromosome (YAC) library is facilitated by information from publicly available databases and published articles. Most numerical chromosome abnormalities, on the other hand, are detrimental to early embryonic development and increase with maternal age. We therefore developed a multichromosome screening technique based on spectral imaging to simultaneously detect and score as many as 10 different chromosome types. The probe set was chosen to detect more than 70% of all numerical chromosome aberrations responsible for spontaneous abortions. Detecting structural and numerical abnormalities in single interphase cells using spectral imaging is a powerful technique for multilocus genetic screening. (J Histochem Cytochem 49:797798, 2001)
Key Words: spectral imaging, chromosome abnormalities, genetic screening, in vitro fertilization, preimplantation genetic, diagnosis
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Introduction |
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BECAUSE CELLS AVAILABLE FOR PGD analysis are likely to be in interphase, we set out to develop rapid procedures for chromosome enumeration and detection of structural abnormalities based on hybridization of chromosome-specific probes and spectral imaging detection. The following describes our typical procedure to prepare case-specific probe for detection of structural chromosome alterations in interphase cells. Institutional Review Board Approval was obtained for this study. Breakpoint-spanning probes were prepared for a carrier of a balanced reciprocal translocation, 46,XY,t(3;4)(p24;p15). We selected YAC clones based on STS markers previously mapped to the approximate breakpoint regions and information provided in the database of the Whitehead Institute for Biomedical Research/MIT Center for Genome Research (
For detection of numerical chromosome abnormalities in interphase cells using spectral imaging, we developed a 10-chromosome probe set (chromosomes 9, 13, 14, 15, 16, 18, 21, 22, X, Y). The chromosomal targets in this set were chosen to detect abnormalities responsible for more than 70% of spontaneous abortions caused by monosomies or trisomies (
In summary, spectral imaging now allows the enumeration of 10 chromosome types in individual interphase cells. Combining the breakpoint-spanning probes with the chromosome enumeration probes will create a very efficient PGD assay for patients carrying balanced translocations. This is expected to significantly increase the success rates in IVF cycles and the chances for affected couples to conceive healthy babies.
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Footnotes |
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Presented in part at the Joint Meeting of the Histochemical Society and the International Society for Analytical and Molecular Morphology, Santa Fe, NM, February 27, 2001.
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Acknowledgments |
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JF was supported in part by an NIEHS training grant (5-T32-ES07106-17).
We wish to thank M. Cassel, J. Garcia, J. Wu, and C. Marquez for skillful assistance, and all anonymous patients who made this study possible by donating embryos.
Received for publication December 5, 2000; accepted February 16, 2001.
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Literature Cited |
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Cassel M, Munné S, Fung J, Weier H-UG (1997) Carrier-specific breakpoint-spanning DNA probes: an approach to preimplantation genetic diagnosis in interphase cells. Hum Reprod 12:2019-2027[Abstract]
Fung J, Hyun W, Dandekar P, Pedersen RA, Weier H-UG (1998a) Spectral imaging in preconception/preimplantation genetic diagnosis (PGD) of aneuploidy: multi-colour, multi-chromosome screening of single cells. J Assist Reprod Genet 15:322-329
Fung J, Munné S, Duell T, Weier H-UG (1998b) Rapid cloning of translocation breakpoints from blood to YAC in 50 days. J Biochem Mol Biol Biophys 1:181-192
Fung J, Weier H-UG, Goldberg JD, Pedersen RA (2000) Multilocus genetic analysis of single interphase cells by spectral imaging. Hum Genet 107:615-622[Medline]
Garini Y, Macville M, du Manoir S, Buckwald RA, Lavi M, Katzir N, Wine D, Bar-Am I, Schröck E, Cabib D, Ried T (1996) Spectral karyotyping. Bioimaging 4:65-72
Hassold TJ, Jacobs PA (1984) Trisomy in man. Annu Rev Genet 18:69-97[Medline]
Hudson TJ, Stein LD, Gerety SS, Ma J, Castle AB, Silva J, Slonim DK, Babtista R, Kruglyak L, Xu SH et al. (1995) An STS-based map of the human genome. Science 270:1945-1954[Abstract]
Munné S, Fung J, Cassel MJ, Márquez C, Weier HUG (1998a) Preimplantation genetic analysis of translocations: case-specific probes for interphase cell analysis. Hum Genet 102:663-674[Medline]
Munné S, Morrison L, Fung J, Márquez C, Weier HUG, BahÁe M, Sable D, Grundfelt L, Schoolcraft B, Scott R, Cohen J (1998b) Spontaneous abortions are reduced after preconception diagnosis of translocations. J Assist Reprod Genet 15:290-296[Medline]
Schröck E, du Manoir S, Veldman T, Schoell B, Wienberg J, FerguesonSmith M, Ning Y, Ledbetter D, Bar-Am I, Soenksen D, Garini Y, Ried T (1996) Multicolor spectral karyotyping of human chromosomes. Science 273:494-497[Abstract]