Skip Navigation


Mol. Hum. Reprod. Advance Access originally published online on August 20, 2004
Molecular Human Reproduction 2004 10(10):767-772; doi:10.1093/molehr/gah101
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
10/10/767    most recent
gah101v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (45)
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Handyside, A. H.
Right arrow Articles by Rutherford, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Handyside, A. H.
Right arrow Articles by Rutherford, A.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Molecular Human Reproduction vol. 10 no. 10 © European Society of Human Reproduction and Embryology 2004; all rights reserved

Isothermal whole genome amplification from single and small numbers of cells: a new era for preimplantation genetic diagnosis of inherited disease

Alan H. Handyside1,2,3,4,*, Mark D. Robinson1,*, Robert J. Simpson3, Mark B. Omar3, Marie-Anne Shaw3, J.Gedis Grudzinskas2 and Anthony Rutherford1

1Leeds PGD Centre, Assisted Conception Unit, Clarendon Wing, Leeds General Infirmary, Leeds, 2The London Bridge Fertility, Gynaecology and Genetics Centre, London and 3School of Biology, University of Leeds, Leeds, UK

4 To whom correspondence should be addressed at: The London Bridge Fertility, Gynaecology and Genetics Centre, 1 St Thomas Street, London Bridge, London SE1 9RY, UK and School of Biology, University of Leeds, Leeds LS2 9JT, UK. Email: a.h.handyside{at}leeds.ac.uk

Preimplantation genetic diagnosis (PGD) of single gene defects following assisted conception typically involves removal of single cells from preimplantation embryos and analysis using highly sensitive PCR amplification methods taking stringent precautions to prevent contamination from foreign or previously amplified DNA. Recently, whole genome amplification has been achieved from small quantities of genomic DNA by isothermal amplification with bacteriophage {phi}29 DNA polymerase- and exonuclease-resistant random hexamer primers. Here we report that isothermal whole genome amplification from single and small numbers of lymphocytes and blastomeres isolated from cleavage stage embryos yielded microgram quantities of amplified DNA, and allowed analysis of 20 different loci, including the {Delta}F508 deletion causing cystic fibrosis and polymorphic repeat sequences used in DNA fingerprinting. As with analysis by PCR-based methods, some preferential amplification or allele drop-out at heterozygous loci was detected with single cells. With 2–5 cells, amplification was more consistent and with 10 or 20 cells results were indistinguishable from genomic DNA. The use of isothermal whole genome amplification as a universal first step marks a new era for PGD since, unlike previous PCR-based methods, sufficient DNA is amplified for diagnosis of any known single gene defect by standard methods and conditions.

Key words: preimplantation genetic diagnosis/single gene defects/whole genome amplification

* Joint first authors


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Hum ReprodHome page
J. Geraedts, J. Collins, L. Gianaroli, V. Goossens, A. Handyside, J. Harper, M. Montag, S. Repping, and A. Schmutzler
What next for preimplantation genetic screening? A polar body approach!
Hum. Reprod., March 1, 2010; 25(3): 575 - 577.
[Abstract] [Full Text] [PDF]

Home page
 Hum ReprodHome page
D.S. Johnson, G. Gemelos, J. Baner, A. Ryan, C. Cinnioglu, M. Banjevic, R. Ross, M. Alper, B. Barrett, J. Frederick, et al.
Preclinical validation of a microarray method for full molecular karyotyping of blastomeres in a 24-h protocol
Hum. Reprod., January 24, 2010; (2010): dep452v1 - dep452.
[Abstract] [Full Text] [PDF]

Home page
 Mol Hum ReprodHome page
J. Ling, G. Zhuang, B. Tazon-Vega, C. Zhang, B. Cao, Z. Rosenwaks, and K. Xu
Evaluation of genome coverage and fidelity of multiple displacement amplification from single cells by SNP array
Mol. Hum. Reprod., November 1, 2009; 15(11): 739 - 747.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
A. Elce, A. Boccia, G. Cardillo, S. Giordano, R. Tomaiuolo, G. Paolella, and G. Castaldo
Three Novel CFTR Polymorphic Repeats Improve Segregation Analysis for Cystic Fibrosis
Clin. Chem., July 1, 2009; 55(7): 1372 - 1379.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
X. Pan, A. E. Urban, D. Palejev, V. Schulz, F. Grubert, Y. Hu, M. Snyder, and S. M. Weissman
A procedure for highly specific, sensitive, and unbiased whole-genome amplification
PNAS, October 7, 2008; 105(40): 15499 - 15504.
[Abstract] [Full Text] [PDF]

Home page
 Am J Trop Med HygHome page
L. J. MORRISON, G. MCCORMACK, L. SWEENEY, A. C.L. LIKEUFACK, P. TRUC, C. M. TURNER, A. TAIT, and A. MACLEOD
USE OF MULTIPLE DISPLACEMENT AMPLIFICATION TO INCREASE THE DETECTION AND GENOTYPING OF TRYPANOSOMA SPECIES SAMPLES IMMOBILIZED ON FTA FILTERS
Am J Trop Med Hyg, June 1, 2007; 76(6): 1132 - 1137.
[Abstract] [Full Text] [PDF]

Home page
 Mol Hum ReprodHome page
Z. Ren, C. Zhou, Y. Xu, J. Deng, H. Zeng, and Y. Zeng
Mutation and haplotype analysis for Duchenne muscular dystrophy by single cell multiple displacement amplification
Mol. Hum. Reprod., June 1, 2007; 13(6): 431 - 436.
[Abstract] [Full Text] [PDF]

Home page
 Mol Hum ReprodHome page
P. Burlet, N. Frydman, N. Gigarel, V. Kerbrat, G. Tachdjian, E. Feyereisen, J.-P. Bonnefont, R. Frydman, A. Munnich, and J. Steffann
Multiple displacement amplification improves PGD for fragile X syndrome
Mol. Hum. Reprod., October 1, 2006; 12(10): 647 - 652.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Z. Jiang, X. Zhang, R. Deka, and L. Jin
Genome amplification of single sperm using multiple displacement amplification
Nucleic Acids Res., June 7, 2005; 33(10): e91 - e91.
[Abstract] [Full Text] [PDF]

Home page
 Mol Hum ReprodHome page
J. F. Sanchez-Garcia, J. Benet, C. Gutierrez-Mateo, J. Luis Seculi, E. Monros, and J. Navarro
Multiple mutation analysis of the cystic fibrosis gene in single cells
Mol. Hum. Reprod., June 1, 2005; 11(6): 463 - 468.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.