Advertisement

FlashFISH: “Same Day” Prenatal Diagnosis of Common Chromosomal Aneuploidies

  • Sherry S. Y. Ho
  • Mahesh A. ChoolaniEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 659)

Abstract

Fluorescence in situ hybridization (FISH) and quantitative fluorescence (QF)-PCR are rapid molecular methods that test for common chromosomal aneuploidies in prenatal diagnosis. While cytogenetic analysis requires approximately 7–14 days before fetal karyotypes are available, these molecular methods release results of sex chromosome aneuploidies, Down syndrome, Edward’s syndrome, and Patau’s syndrome within 24–48 h of fetal sampling, alleviating parental anxiety. However, specific diagnosis or exclusion of aneuploidy should be available within the same day of amniocentesis. We developed “FlashFISH,” a low cost FISH method that allows accurate results to be reported within 2 h of fetal sampling. Here, we report our experience of using FlashFISH in prenatal diagnosis, and we illustrate in detail the protocols used for the purpose in our laboratory.

Key words

Cytogenetics Down syndrome FISH Molecular diagnosis Prenatal diagnosis 

References

  1. 1.
    Evans, M. I., Henry, G. P., Miller, W. A., Bui, T. H., Snidjers, R. J., Wapner, R. J., Miny, P., Johnson, M. P., Peakman, D., Johnson, A., Nicolaides, K., Holzgreve, W., Ebrahim, S. A., Babu, R., Jackson, L. (1999) International, collaborative assessment of 146,000 prenatal karyotypes: Expected limitations if only chromosome-specific probes and fluorescent in-situ hybridization are used. Hum. Reprod. 14, 1213–1216.PubMedCrossRefGoogle Scholar
  2. 2.
    Caine, A., Maltby, A. E., Parkin, C. A., Waters, J. J., Crolla, J. A. (2005) Prenatal detection of Down’s syndrome by rapid aneuploidy testing for chromosomes 13, 18, and 21 by FISH or PCR without a full karyotype: A cytogenetic risk assessment. Lancet 366, 123–128.PubMedCrossRefGoogle Scholar
  3. 3.
    Klinger, K., Landes, G., Shook, D., Harvey, R., Lopez, L., Locke, P., Lerner, T., Osathanondh, R., Leverone, B., Houseal, T., et al. (1992) Rapid detection of chromosome aneuploidies in uncultured amniocytes by using fluorescence in situ hybridization (FISH). Am. J. Hum. Genet. 51, 55–65.PubMedGoogle Scholar
  4. 4.
    Weise, A., Liehr, T. (2008) Rapid prenatal aneuploidy screening by fluorescence in situ hybridization (FISH). Methods Mol. Biol. 444, 39–47.PubMedCrossRefGoogle Scholar
  5. 5.
    Pertl, B., Yau, S. C., Sherlock, J., Davies, A. F., Mathew, C. G., Adinolfi, M. (1994) Rapid molecular method for prenatal detection of Down’s syndrome. Lancet 343, 1197–1198.PubMedCrossRefGoogle Scholar
  6. 6.
    Mann, K., Petek, E., Pertl, B. (2008) Prenatal detection of chromosome aneuploidy by quantitative fluorescence PCR. Methods Mol. Biol. 444, 71–94.PubMedCrossRefGoogle Scholar
  7. 7.
    Schouten, J. P., McElgunn, C. J., Waaijer, R., Zwijnendburg, D., Diepvens, F., Pals, G. (2002) Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. Nucleic Acids Res. 30, e57.PubMedCrossRefGoogle Scholar
  8. 8.
    Kooper, A. J., Faas, B. H., Kater-Baats, E., Feuth, T., Janssen, J. C., van der Burgt, I., Lotgering, F. K., Geurts van Kessel, A., Smits, A. P. (2008) Multiplex Ligation-dependent Probe Amplification (MLPA) as a standalone test for rapid aneuploidy detection in amniotic fluid cells. Prenat. Diagn. 28, 1004–1010.PubMedCrossRefGoogle Scholar
  9. 9.
    Tercyak, K. P., Johnson, S. B., Roberts, S. F., Cruz, A. C. (2001) Psychological response to prenatal genetic counseling and amniocentesis. Patient Educ. Couns. 43, 73–84.PubMedCrossRefGoogle Scholar
  10. 10.
    Jouannic, J. M., Martinovic, J., Bessières, B., Romana, S., Bonnet, D. (2003) Fluorescence in situ hybridization (FISH) rather than ultrasound for the evaluation of fetuses at risk for 22q11.1 deletion. Prenat. Diagn. 23, 607–608.PubMedCrossRefGoogle Scholar
  11. 11.
    Shaffer, L. G., Bui, T-H. (2007) Molecular cytogenetic and rapid aneuploidy detection methods in prenatal diagnosis. Am. J. Med. Genet. C. Semin. Med. Genet. 145C, 87–98.PubMedCrossRefGoogle Scholar
  12. 12.
    Choolani, M., Ho, S. S., Razvi, K., Ponnusamy, S., Baig, S., Fisk, N. M., Biswas, A., Rapid Molecular Testing in Prenatal Diagnosis Group. (2007) FastFISH: Technique for ultrarapid fluorescence in situ hybridization on uncultured amniocytes yielding results within 2 h of amniocentesis. Mol. Hum. Reprod. 13, 355–359.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of Obstetrics and Gynaecology, Yong Loo Lin School of MedicineNational University of SingaporeSingaporeSingapore

Personalised recommendations