Development of ultrasensitive diagnostic assays for the detection of genetic disorders, infections and for monitoring the therapeutic success following gene therapy

• Unité de recherche: PCRD EU
• Numéro de projet: 98.0264
• Titre du projet anglais: Development of ultrasensitive diagnostic assays for the detection of genetic disorders, infections and for monitoring the therapeutic success following gene therapy

Mots-clé (Anglais)

Molecular diagnostics; quantitative PCR; FRET

Autre Numéro de projet (Anglais)

EU project number: BMH4-CT98-9579

Programme de recherche (Anglais)

EU-programme: 4. Frame Research Programme - 4.2 Agriculture and agroindustry

Description succincte (Anglais)

See abstract

Partenaires et organisations internationales (Anglais)

Coordinator: Paul-Ehrlich-Institut, Langen (D)

Résumé des résultats (Abstract) (Anglais)

The major objective of this project is to provide a novel highly sensitive PCR based detection method for both, the molecular diagnosis and the therapeutic monitoring of diseases which are related to genetic disorders and infections. The principle of fluorescence resonance energy transfer (FRET) as the backbone technology was adapted for monitoring the PCR reaction. Extremely low amounts of DNA in the ng scale have been detected with the potential of real time PCR (multi endpoint determination). Specific gene targets were identified and primers were designed and coupled to different dyes for efficient application in FRET based technology. Texas Red and Fluorescine have been selected by intense testing being the suitable dyes for our PCR based diagnostic system. The following tasks have been executed by Partners within the second period of funding and appropriate milestones have been achieved, 1.) optimisation of wet chemical components and establishment of diagnostic assay protocols for laboratory usag 2.) establishment of a 'tail sequence' and coupling reagent, 3.) design of hardware stage II, 4.) Modular design of Software and subsequent adaptation to instrumentation control.

Références bases de données (Anglais)

Swiss Database: Euro-DB of the
State Secretariat for Education and Research
Hallwylstrasse 4
CH-3003 Berne, Switzerland
Tel. [phone]
Swiss Project-Number: 98.0264