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Q01085: The quantitative determination of common wheat in durum pasta using real-time PCR method-validation

Monday 25 October 2004

This research project aims to validate a DNA method to quantify the amount of common wheat present in durum wheat pasta.

Study Duration : June 2003 to February 2004

Contractor : RHM Technology Ltd

Background

The authenticity of pasta made from durum wheat ( Triticum durum ) and the detection and quantification of deliberate adulteration of T.durum pastas by common wheat ( T. aestivum ) continues to be an important issue throughout Europe.

Methodologies currently available have a number of shortcomings, which limit their ability to accurately quantify the level of common wheat in Durum pasta. A promising real-time polymerase chain reaction (PCR) method to quantify common wheat in durum pasta is based on amplification of a small sequence of DNA on the D-genome of common wheat. Before this method can be used by the Agency or by enforcement officers it must be fully validated and a standard operating protocol (SOP) prepared. The aim of this project is to establish the performance criteria (sensitivity, accuracy, precision) of the developed method, and to determine its suitability for use in a survey of pasta products.

Research Approach

In addition to establishing the method performance criteria, a key part of the project is to create suitable standard reference materials with which to calibrate analysis. A comparison of existing pasta reference materials and flour blends will be made to confirm that the method used is insensitive to processing history. If successful, this would allow the cheaper production of reference materials. A performance standard containing 3% common wheat ( T.aestivum ) flour will be made with the aim of enabling a quantitative control to be included in all analytical work and to allow an assessment of uncertainty to be made. The accuracy of the method will also be established, prior to preparation of an SOP.

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