Protocol proposal for Friedreich ataxia molecular diagnosis using fluorescent and triplet repeat primed polymerase chain reaction

Transl Res. 2010 Nov;156(5):309-14. doi: 10.1016/j.trsl.2010.08.001.


Friedreich ataxia (FRDA) is the most common hereditary ataxia that is caused mainly by an unstable GAA trinucleotide expansion in the first intron of the frataxin gene. Molecular tests for FRDA diagnosis and carrier detection include polymerase chain reaction (PCR) for the GAA expansion, triplet repeat primed PCR (TP-PCR), and/or Southern blotting. TP-PCR is a method developed to detect trinucleotide expansions successfully applied to FRDA diagnosis. In our laboratory, we have included a PCR for the GAA expansion using fluorescent primers polymerase chain reaction (F-PCR) to identify normal heterozygous and affected individuals unambiguously. The purpose of our study was to reanalyze 310 samples previously diagnosed in our laboratory and compare the results with those obtained by F-PCR and TP-PCR. Eight percent of the discrepancies between the carrier and the normal individuals were identified correctly by this protocol. No discrepancy was detected in the affected individuals. These techniques are effective, and compared with Southern blotting, they are less labor-intensive and suitable for automation. We suggest a new routine protocol for FRDA diagnosis that includes F-PCR and TP-PCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Clinical Protocols
  • DNA Mutational Analysis
  • Friedreich Ataxia / diagnosis*
  • Friedreich Ataxia / genetics
  • Humans
  • Molecular Diagnostic Techniques
  • Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Research Design
  • Trinucleotide Repeat Expansion / genetics