Document Detail

The minisequencing method: an alternative strategy for preimplantation genetic diagnosis of single gene disorders.
MedLine Citation:
PMID:  12802047     Owner:  NLM     Status:  In-Process    
We have applied a new method of genetic analysis, called 'minisequencing', to preimplantation genetic diagnosis (PGD) of monogenic disorders from single cells. This method involves computer-assisted mutation analysis, which allows exact base identity determination and computer-assisted visualization of the specific mutation(s), and thus facilitates data interpretation and management. Sequencing of the entire PCR product is unnecessary, yet the same qualitative characteristics of sequence analysis are maintained. The main benefit of the minisequencing strategy is the use of a mutation analysis protocol based on a common procedure, irrespective of the mutations involved. To evaluate the reliability of this method for subsequent application to PGD, we analysed PCR products from 887 blastomeres including 55 PGD cases of different genetic diseases, such as cystic fibrosis, beta-thalassaemia, sickle cell anaemia, haemophilia A, retinoblastoma, and spinal muscular atrophy. Minisequencing was found to be a useful technique in PGD analysis, due to its elevated sensitivity, automation, and easy data interpretation. The method was also efficient, providing interpretable results in 96.5% (856/887) of the blastomeres tested. Fifteen clinical pregnancies resulted from these PGD cases; conventional prenatal diagnosis confirmed all the PGD results, and 10 healthy babies have already been born. Its applicability to PGD could be helpful, particularly in cases in which the mutation(s) involved are difficult to assess by restriction analysis or other commonly used methods.
F Fiorentino; M C Magli; D Podini; A P Ferraretti; A Nuccitelli; N Vitale; M Baldi; L Gianaroli
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Molecular human reproduction     Volume:  9     ISSN:  1360-9947     ISO Abbreviation:  Mol. Hum. Reprod.     Publication Date:  2003 Jul 
Date Detail:
Created Date:  2003-06-12     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9513710     Medline TA:  Mol Hum Reprod     Country:  England    
Other Details:
Languages:  eng     Pagination:  399-410     Citation Subset:  IM    
'Genoma' Molecular Genetics Laboratory, via Po nr 102, 00198 Rome.
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