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Clinical Chemistry 42: 1601-1603, 1996;
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Clinical Chemistry, Vol 42, 1601-1603, Copyright © 1996 by American Association for Clinical Chemistry

Nonisotopic method for accurate detection of (CAG)n repeats causing Huntington disease

M Muglia, O Leone, G Annesi, AL Gabriele, E Imbrogno, C Grandinetti, FL Conforti, F Naso and C Brancati
Istituto di Medicina Sperimentale e Biotecnologie, CNR, Cosenza, Italy. Muglia@Area.CS.CNR.IT

Huntington disease (HD) is a neurodegenerative disorder caused by an expanded trinucleotide repeat (CAG)n located at the 5' end of the novel IT15 gene. Discovery of this expansion allows the molecular diagnosis of HD by measuring repeat length. We applied a simple nonisotopic method to detect (CAG)n repeats, avoiding both radioactive and Southern transfer analysis. The assay is based on direct visualization of electrophoresed PCR products, after silver nitrate gel staining. Its accurate sizing of HD alleles allows presymptomatic diagnosis of at- risk persons. By avoiding isotopic manipulations, the method is safe and accurate, with no radioactive background bands. Furthermore, because it permits direct allele visualization after gel staining, the method is simple and rapid, allowing allele sizing within hours rather than days.


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