Short Communication: Rapid Detection of the 
F508 Mutation in Single Cells Using DHPLC: Implications for Preimplantation Genetic Diagnosis
Authors: Girardet A.1; Cathala P.2; Claustres M.2
Source: Journal of Assisted Reproduction and Genetics, Volume 20, Number 4, April 2003 , pp. 153-156(4)
Publisher: Springer
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Abstract:
Purpose: Practice of preimplantation genetic diagnosis (PGD) requires efficient amplification and analysis techniques. We have tested Denaturing High Performance Liquid Chromatography (DHPLC) to screen the F508 mutation in heterozygous single cells in order to assess its usefulness for PGD of cystic fibrosis.
Methods: One hundred and two single lymphocytes—including N/N and N/F508—were studied. F508 locus was amplified by nested PCR followed by the analysis of PCR products by DHPLC in non-denaturing conditions.
Results: On the basis of PCR-amplified product analysis, total efficiency of amplification was 98.78% (101/102), and allele dropout (ADO) rate was 3.7% (3/81). For each sample, results were obtained in less than 4 min with high resolution.
Conclusions: DHPLC is a rapid and efficient technique to detect the F508 mutation in single cells and is therefore appropriate for clinical application of preimplantation genetic diagnosis of cystic fibrosis.
Keywords:
Cystic fibrosis;
DHPLC;
F508;
preimplantation genetic diagnosis
Language: English
Document Type: Short communication
Affiliations: 1: Laboratoire de Génétique Moléculaire, Centre Hospitalo-Universitaire (CHU) and Institut Universitaire de Recherche Clinique (IURC), 641 Avenue du Doyen Gaston Giraud, 34093 Montpellier cedex 5, France; girardet@igh.cnrs.fr 2: Laboratoire de Génétique Moléculaire, Centre Hospitalo-Universitaire (CHU) and Institut Universitaire de Recherche Clinique (IURC), 641 Avenue du Doyen Gaston Giraud, 34093 Montpellier cedex 5, France
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