Naslov
Expand long PCR for fragile X mutation detection

Autori
Hećimović, Silva ; Barišić, Ingeborg ; Ligutić, Ivo ; Pavelić, Krešimir

Izvornik
Medizinische Genetik (0936-5931) 9 (1997), 2;

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, kongresno priopcenje, znanstveni

Ključne riječi
Expand long PCR; fragile X syndrome

Sažetak
Fragile X mutation detection by DNA analysis has enabled accurate diagnosis of the fragile X syndrome. The mutation, which involves the expansion of CGG repeats in the FMR1 gene, has been primarily detected by Southern blotting method. In this study we present a novel, efficient and reliable PCR protocol that is more convenient for routine diagnosis of the fragile X syndrome. This method is based on the use of Expand Long PCR System which enables the amplification of normal, premutated and full mutated alleles, and therefore provides a complete CGG repeat analysis of the FMR1 gene. Normal alleles were easily detected by ethidium bromide staining of the agarose gels, suggesting that this assay could be used as a screening test for a large number of referrals. The amplified premutations and full mutations were identified by hybridisation with a digoxigenin labelled 5´-(CGG)5-3´ probe, followed by chemiluminescent detection. In this study we propose a new strategy for molecular diagnosis of the fragile X sydnorme in which our Expand Long PCR protocol should be used as the first screening test for fragile X mutation detection.

Izvorni jezik
Engleski

Znanstvena područja
Kliničke medicinske znanosti

POVEZANOST RADA