Molecular Human Reproduction, Vol. 5, No. 12, 1166-1175,
December 1999
© 1999 European Society of Human Reproduction and Embryology
Diagnosing genetic disease |
Assessment of multiplex fluorescent PCR for screening single cells for trisomy 21 and single gene defects
1 Department of Obstetrics and Gynecology, and 2 Department of Human Genetics, Royal Victoria Hospital, McGill University, Montreal, Canada
Abstract
A great majority of patients seeking preimplantation genetic diagnosis (PGD) are women >35 years of age. In addition to being carriers for single gene defects, these women also have a higher risk of having children with Down's syndrome (trisomy 21). For these patients, it would be advantageous if a diagnostic test for trisomy 21 was developed, which could be used in conjunction with tests for single gene defects. Here, we assessed the feasibility of developing an accurate genetic test for diagnosing trisomy 21 and the mutation causing spinal muscular atrophy (SMA) in single cells using multiplex fluorescence polymerase chain reaction (PCR). Single- and two-round PCR were developed using a combination of primers for the survival motor neuron (SMN) gene exons 7 and 8 and two chromosome 21 short tandem repeats (STRs), D21S226 and D21S11. After only 36 cycles, 88 and 68% of normal single cells were screened for SMA mutations and trisomy 21 respectively. In multiplex PCR using only two primers (SMN exon 7 and D21S11) instead of four, the efficiency of SMA diagnosis was increased to 93%. In the same reactions, the D21S11 alleles were detected in 83% of the normal single cells. Clinical applications of this assay should enable detection of those embryos that have inherited three heterozygous alleles and, therefore, benefit many PGD patients who are at an increased risk of Down's syndrome.
multiplex fluorescent PCR/single cell/spinal muscular atrophy/trisomy 21
Notes
3 To whom correspondence should be addressed
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  F. Fiorentino, M.C. Magli, D. Podini, A.P. Ferraretti, A. Nuccitelli, N. Vitale, M. Baldi, and L. Gianaroli The minisequencing method: an alternative strategy for preimplantation genetic diagnosis of single gene disorders Mol. Hum. Reprod., July 1, 2003; 9(7): 399 - 410. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M.G. Katz, A.O. Trounson, and D.S. Cram DNA fingerprinting of sister blastomeres from human IVF embryos Hum. Reprod., March 1, 2002; 17(3): 752 - 759. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. R. Thornhill and K. Snow Molecular Diagnostics in Preimplantation Genetic Diagnosis J. Mol. Diagn., February 1, 2002; 4(1): 11 - 29. [Full Text] [PDF]
|
|
|
|
|
|
G. Daniels, R. Pettigrew, A. Thornhill, S. Abbs, A. Lashwood, F. O'Mahony, C. Mathew, A. Handyside, and P. Braude Six unaffected livebirths following preimplantation diagnosis for spinal muscular atrophy Mol. Hum. Reprod., October 1, 2001; 7(10): 995 - 1000. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N.L. Dean, S.L. Tan, and A. Ao The development of preimplantation genetic diagnosis for myotonic dystrophy using multiplex fluorescent polymerase chain reaction and its clinical application Mol. Hum. Reprod., September 1, 2001; 7(9): 895 - 901. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Samura, S. Sohda, K. L. Johnson, B. Pertl, S. Ralston, L. C. Delli-Bovi, and D. W. Bianchi Diagnosis of Trisomy 21 in Fetal Nucleated Erythrocytes from Maternal Blood by Use of Short Tandem Repeat Sequences Clin. Chem., September 1, 2001; 47(9): 1622 - 1626. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.C.F.M. Dreesen, L.J.A.M. Jacobs, M. Bras, J. Herbergs, J.C.M. Dumoulin, J.P.M. Geraedts, J.L.H. Evers, and H.J.M. Smeets Multiplex PCR of polymorphic markers flanking the CFTR gene; a general approach for preimplantation genetic diagnosis of cystic fibrosis Mol. Hum. Reprod., May 1, 2000; 6(5): 391 - 396. [Abstract] [Full Text] [PDF]
|
|