An enhanced polymerase chain reaction assay to detect pre- and full mutation alleles of the fragile X mental retardation 1 gene

Alessandro Saluto, Alessandro Brussino, Flora Tassone, Carlo Arduino, Claudia Cagnoli, Patrizia Pappi, Paul J Hagerman, Nicola Migone, Alfredo Brusco

Research output: Contribution to journalArticle

86 Scopus citations

Abstract

Several diagnostic strategies have been applied to the detection of FMR1 gene repeat expansions in fragile X syndrome. Here, we report a novel polymerase chain reaction-based strategy using the Expand Long Template PCR System (Roche Diagnostics, Mannheim, Germany) and the osmolyte betaine. Repeat expansions up to ∼330 CGGs in males and up to at least ∼160 CGGs in carrier women could be easily visualized on ethidium bromide agarose gels. We also demonstrated that fluorescence analysis of polymerase chain reaction products was a reliable tool to verify the presence of premutation and full mutation alleles both in males and in females. This technique, primarily designed to detect premutation alleles, can be used as a routine first screen for expanded FMR1 alleles.

Original languageEnglish (US)
Pages (from-to)605-612
Number of pages8
JournalJournal of Molecular Diagnostics
Volume7
Issue number5
StatePublished - Nov 2005

ASJC Scopus subject areas

  • Molecular Biology

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