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Research Project to Validate Methods to Quantify Horse and Pork Meat Adulteration and Processed Beef - FA0171

The horsemeat crisis lead to a concerted FSA/Defra response that resulted in the development and subsequent validation of a qPCR method to quantitate horse meat in a background of raw beef (meat) to a 1% (w/w) level, in line with EU Commission protocol for the threshold for deliberate vs. adventitious adulteration. The protocol was a pragmatic threshold based on a project undertaken by the EURL-AP following the horsemeat discovery, which showed proof of principle that a semi-quantitative approach (such as real-time qPCR) can establish a 1% adulteration of horse meat in another meat within a 95% confidence interval.

A proof of principle project (FA0134) showed that a quantitative PCR based-approach on two methods in the literature was applicable to quantifying horsemeat in a background of beef (raw: processed and unprocessed). The following project, FA0135, developed the horse qPCR method, and was subsequently validated in-house in Defra-project FA0146. The FSA funded an international ring-trial (FSA 126001) of the method to further validate it for use by public analysts for food law enforcement.

A follow up research project (FA0157) compared ‘real-time’ qPCR with a digital PCR (dPCR) method. The research project aimed to assess proof of principle that a qPCR method could be applied to (cooked) processed beef samples, as well as if it could be applied to two novel quantitation assays: pork-in-beef and beef-in-lamb. The pork-in-beef assay is of particular importance due to the cultural implications of mislabelling of pork for faith groups. The results indicated that the real-time qPCR and dPCR method is applicable for both the horse-in-beef and the pork-in-beef assays for both raw and processed (cooked) samples e.g. a cottage pie ready meal. Despite this, it was advised by our expert technical group that digital PCR should not be followed up on at this current time due to the its relative expense compared to qPCR, as well as the lack of dPCR capabilities in Public Analyst labs.

Therefore, the purpose of this research is to build on what has been developed so far to extend the use of real-time qPCR pork and horsemeat adulteration in processed (cooked) beef matrices e.g. cottage pie. The Authenticity Methods Working Group (AMWG) has highlighted this research need based on intelligence that meat substitution is still likely to be the most common manifestation of food fraud experienced by UK consumers, coupled with the cultural importance of diversifying methods to apply to pork testing.
Objective 1 – Single lab (intra-lab) validation of the horse-in-beef and pork-in-beef qPCR assays

Objective 2 – Inter-laboratory validation of the horse-in-beef and pork-in-beef qPCR assays

Project Documents
• EVID4 - Final project report : Defra FA0171 EVID4 Final Report   (620k)
• ANX - Annex : Annex 1 - Draft SOPs v3   (8414k)
Time-Scale and Cost
From: 2018

To: 2019

Cost: £114,381
Contractor / Funded Organisations
LGC Limited
Food Chain              
Meat Quality