Rapid Detection of the ΔF508 Mutation in Single Cells Using DHPLC: Implications for Preimplantation Genetic Diagnosis.

Autor: Girardet, Anne, Cathala, Philippe, Claustres, Mireille
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Zdroj: Journal of Assisted Reproduction & Genetics; Apr2003, Vol. 20 Issue 4, p153, 4p, 2 Diagrams, 1 Chart
Abstrakt: 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 ΔF508 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/ΔF508—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 ΔF508 mutation in single cells and is therefore appropriate for clinical application of preimplantation genetic diagnosis of cystic fibrosis. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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