Molecular Human Reproduction, Vol 3, 523-528, Copyright © 1997 by Oxford University Press
E Kanavakis, J Traeger-Synodinos, C Vrettou, E Maragoudaki, M Tzetis and C Kattamis
Prenatal diagnostic strategies applied today are based mainly on polymerase
chain reaction (PCR) analytical protocols. In Greece a wide range of
mutations underlie the thalassaemic haemoglobinopathies, and consequently a
variety of PCR-based methods are required to facilitate diagnosis of all
potential abnormal genotypes. PCR protocols include those which are
relatively simple and others that are technically challenging, but very few
have been designed for high through-put clinical diagnostics. Over a period
of 18 months we carried out prenatal diagnosis of 147 pregnancies (150
fetal samples) at risk for a wide range of haemoglobinopathies. This
involved the precise characterization of parental genotypes and the
subsequent analysis of fetal DNA samples. In this series, 18 different
mutations in the alpha- or beta-globin clusters were identified. For the
characterization of these mutations, five PCR-based protocols were
selected: denaturing gradient gel electrophoresis (DGGE), amplification
refractory mutation system (ARMS) PCR, restriction endonuclease analysis of
PCR fragments, oligonucleotide hybridization and 'gap' PCR for detection of
deletions. To avoid spurious diagnosis due to contamination of fetal
samples, two additional methods were used to genotype polymorphic variable
nucleotide tandem repeat (VNTR) regions of the genome in parental and fetal
samples. Through analysis of the results we assess the advantages and
drawbacks of the selected PCR-based protocols for providing routine
clinical diagnostics.
JOURNAL ARTICLE
Prenatal diagnosis of the thalassaemia syndromes by rapid DNA analytical methods
First Department of Pediatrics, Athens University, St. Sophia's Children's Hospital, Greece.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  K. Glynou, P. Kastanis, S. Boukouvala, V. Tsaoussis, P. C. Ioannou, T. K. Christopoulos, J. Traeger-Synodinos, and E. Kanavakis High-Throughput Microtiter Well-Based Chemiluminometric Genotyping of 15 HBB Gene Mutations in a Dry-Reagent Format Clin. Chem., March 1, 2007; 53(3): 384 - 391. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Traeger-Synodinos, C. Vrettou, G. Palmer, M. Tzetis, M. Mastrominas, S. Davies, and E. Kanavakis An evaluation of PGD in clinical genetic services through 3 years application for prevention of {beta}-thalassaemia major and sickle cell thalassaemia Mol. Hum. Reprod., May 1, 2003; 9(5): 301 - 307. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Vrettou, J. Traeger-Synodinos, M. Tzetis, G. Malamis, and E. Kanavakis Rapid Screening of Multiple {beta}-Globin Gene Mutations by Real-Time PCR on the LightCycler: Application to Carrier Screening and Prenatal Diagnosis of Thalassemia Syndromes Clin. Chem., May 1, 2003; 49(5): 769 - 776. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Vrettou, M. Tzetis, J. Traeger-Synodinos, G. Palmer, and E. Kanavakis Multiplex sequence variation detection throughout the CFTR gene appropriate for preimplantation genetic diagnosis in populations with heterogeneity of cystic fibrosis mutations Mol. Hum. Reprod., September 1, 2002; 8(9): 880 - 886. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G.A. Palmer, J. Traeger-Synodinos, S. Davies, M. Tzetis, C. Vrettou, M. Mastrominas, and E. Kanavakis Pregnancies following blastocyst stage transfer in PGD cycles at risk for {beta}-thalassaemic haemoglobinopathies Hum. Reprod., January 1, 2002; 17(1): 25 - 31. [Abstract] [Full Text] [PDF]
|
|