Different approaches for noninvasive prenatal diagnosis of genetic diseases based on PNA-mediated enriched PCR

Silvia Galbiati, Gabriella Restagno, Barbara Foglieni, Sara Bonalumi, Maurizio Travi, Antonio Piga, Luca Sbaiz, Marcella Chiari, Francesco Damin, Maddalena Smid, Luca Valsecchi, Federica Pasi, Augusto Ferrari, Maurizio Ferrari, Laura Cremonesi

Research output: Chapter in Book/Report/Conference proceedingConference contribution


The aim of this work was to develop advanced and accessible protocols for noninvasive prenatal diagnosis of genetic diseases. We are evaluating different technologies for mutation detection, based on fluorescent probe hybridization of the amplified product and pyrosequencing, a technique that relies on the incorporation of nucleotides in a primer-directed polymerase extension reaction. In a previous investigation, we have already proven that these approaches are sufficiently sensitive to detect a few copies of a minority-mutated allele in the presence of an excess of wild-type DNA, In this work, in order to further enhance the sensitivity, we have employed a mutant enrichment amplification strategy based on the use of peptide nucleic acids (PNAs). These DNA analogues bind wild-type DNA, thus interfering with its amplification while still allowing the mutant DNA to become detectable. We have synthesized different PNAs, which are highly effective in clamping wild-type DNA in the beta-globin gene region, where four beta-thalassemia mutations are located (IVSI.110, CD39, IVSI.1, IVSI.6) plus HbS. The fluorescence microchip readout allows us to monitor the extent of wild-type allele inhibition, thus facilitating the assessment of the optimal PNA concentration.

Original languageEnglish
Title of host publicationAnnals of the New York Academy of Sciences
Number of pages7
Publication statusPublished - Sep 2006

Publication series

NameAnnals of the New York Academy of Sciences
ISSN (Print)00778923
ISSN (Electronic)17496632


  • Beta-thalassemia
  • Fetal DNA in maternal plasma
  • Noninvasive prenatal diagnosis
  • PNA-mediated enriched PCR

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


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