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 journalArticle

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

Fingerprint

Craniosynostoses
Denaturation
Gene Amplification
Carbon Monoxide
Prenatal Diagnosis
Amplification
Genes
Alleles
Polymerase Chain Reaction
Inborn Genetic Diseases
Temperature
Mothers
Chorionic Villi
Mutation
Plasmas
DNA

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)

Cite this

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

In: Clinical Chemistry and Laboratory Medicine, Vol. 52, No. 4, 01.04.2014, p. 505-509.

Research output: Contribution to journalArticle

@article{6f1faedd69a24671a7586698f6767600,
title = "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",
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.",
keywords = "craniosynostosis, free fetal DNA in maternal plasma, full COLD-PCR, genetic diseases, non-invasive prenatal diagnosis",
author = "Silvia Galbiati and Stefania Stenirri and Luca Sbaiz and Marco Barberis and Laura Cremonesi and Gabriella Restagno and Maurizio Ferrari",
year = "2014",
month = "4",
day = "1",
doi = "10.1515/cclm-2013-0757",
language = "English",
volume = "52",
pages = "505--509",
journal = "Clinical Chemistry and Laboratory Medicine",
issn = "1434-6621",
publisher = "Walter de Gruyter GmbH",
number = "4",

}

TY - JOUR

T1 - 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

T2 - First case report

AU - Galbiati, Silvia

AU - Stenirri, Stefania

AU - Sbaiz, Luca

AU - Barberis, Marco

AU - Cremonesi, Laura

AU - Restagno, Gabriella

AU - Ferrari, Maurizio

PY - 2014/4/1

Y1 - 2014/4/1

N2 - 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.

AB - 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.

KW - craniosynostosis

KW - free fetal DNA in maternal plasma

KW - full COLD-PCR

KW - genetic diseases

KW - non-invasive prenatal diagnosis

UR - http://www.scopus.com/inward/record.url?scp=84896793710&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84896793710&partnerID=8YFLogxK

U2 - 10.1515/cclm-2013-0757

DO - 10.1515/cclm-2013-0757

M3 - Article

C2 - 24166674

AN - SCOPUS:84896793710

VL - 52

SP - 505

EP - 509

JO - Clinical Chemistry and Laboratory Medicine

JF - Clinical Chemistry and Laboratory Medicine

SN - 1434-6621

IS - 4

ER -