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

• Research unit: EU RFP
• Project number: 98.0264

Key words (English)

Molecular diagnostics; quantitative PCR; FRET

Alternative project number (English)

EU project number: BMH4-CT98-9579

Research programs (English)

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

Short description (English)

See abstract

Partners and International Organizations (English)

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

Abstract (English)

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.

References in databases (English)

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