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Mol. Hum. Reprod. Advance Access originally published online on October 1, 2004
Molecular Human Reproduction 2004 10(11):847-852; doi:10.1093/molehr/gah114
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Molecular Human Reproduction vol. 10 no. 11 © European Society of Human Reproduction and Embryology 2004; all rights reserved

Multiple displacement amplification on single cell and possible PGD applications

Ali Hellani1,3, Serdar Coskun1, Moncef Benkhalifa2, Abelghani Tbakhi1, Nadia Sakati1, Ali Al-Odaib1 and Pinar Ozand1

1King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia and 2ATL R&D Laboratory, Paris, France

3 To whom correspondence should be addressed at: King Faisal Specialist Hospital and Research Center, P.O.Box 3354 MBC #10 Riyadh, 11211 Saudi Arabia. Email: hellani{at}kfshrc.edu.sa

Multiple displacement amplification (MDA) is a technique used in the amplification of very low amounts of DNA and reported to yield large quantities of high-quality DNA. We used MDA to amplify the whole genome directly from a single cell. The most common techniques used in PGD are PCR and fluorescent in-situ hybridization (FISH). There are many limitations to these techniques including, the number of chromosomes diagnosed for FISH or the quality of DNA issued from a single cell PCR. This report shows, for the first time, use of MDA for single cell whole genome amplification. A total of 16 short tandem repeats (STRs) were amplified successfully with a similar pattern to the genomic DNA. Furthermore, allelic drop out (ADO) derived from MDA was assessed in 40 single cells by analysing (i) heterozygosity for a known ß globin mutation (IVSI-5 C–G) and by studying (ii) the heterozygous loci present in the STRs. ADO turned out to be 10.25% for the ß globin gene sequencing and 5% for the fluorescent PCR analysis of STRs. Moreover, the amplification accuracy of MDA permitted the detection of trisomy 21 on a single cell using comparative genome hybridization-array. Altogether, these data suggest that MDA can be used for single cell molecular karyotyping and the diagnosis of any single gene disorder in PGD.

Key words: PGD/whole genome amplification-comparative genome hybridization/CGH-array/multiple displacement amplification


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