Identification of an 18 bp deletion in the TWIST1 gene by CO-amplification at lower denaturation temperature-PCR (COLD-PCR) for non-invasive prenatal diagnosis of craniosynostosis: First case report

Silvia Galbiati, Stefania Stenirri, Luca Sbaiz, Marco Barberis, Laura Cremonesi, Gabriella Restagno, Maurizio Ferrari

Research output: Contribution to journalArticlepeer-review

Abstract

Background: Non-invasive prenatal diagnosis has found application in a limited number of genetic diseases due to the difficulty in detecting a few copies of fetal mutated sequences in the presence of a large excess of wild-type maternal alleles, even in the case of single-base mutations. Methods: We developed conditions for the enrichment of fetal mutated alleles in maternal plasma based on COamplification at lower denaturation temperature-PCR (COLD-PCR). In particular, we applied a full COLD-PCR protocol to the identification of a p.A87-G92del mutation in the TWIST1 gene causing craniosynostosis in a couple at risk for the disease. Results: The use of the COLD-PCR protocol coupled with direct sequencing enabled correct identification of the fetal paternally inherited mutated allele, in accordance with the result obtained on DNA extracted from chorionic villi. Conclusions: COLD-PCR proved to be a simple and powerful tool for the identification of minority mutated alleles even in the case of a moderately large deletion (18 bp) and confirmed to be very suitable for non-invasive prenatal diagnosis of a variety of genetic diseases.

Original languageEnglish
Pages (from-to)505-509
Number of pages5
JournalClinical Chemistry and Laboratory Medicine
Volume52
Issue number4
DOIs
Publication statusPublished - Apr 1 2014

Keywords

  • craniosynostosis
  • free fetal DNA in maternal plasma
  • full COLD-PCR
  • genetic diseases
  • non-invasive prenatal diagnosis

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical
  • Medicine(all)

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