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Detection & identification of species-specific nucleic acids in rendered animal material - SE1755

In 1996 MAFF introduced a complete ban of mammalian meat and bonemal from all animal feed. To ensure that companies comply with this ruling specific methods are required that can detect and differentiate between the proteins of different species. This project aims to develop such an assay to detect and differentiate between cattle, sheep, pig and chicken proteins, based on the detection of nucleic acids. A PCR assay could be used as stand-alone screening assay or act as a backup, confirmatory test of existing immunoassays.
01. Determine the thermostability of DNA by heating tissues in the range 90-133 degrees C at 3 bar for 20 minutes.
02. Establish DNA extraction protocol for raw and rendered material (different processing procedures).
03. Design PrP, albumin or mitochondrial DNA and actin amplification primer pairs and potential fluorescent probes for all species in this study.
04. Assess the various amplification primers and probes on DNA extracted from raw tissues for all species. Determine PCR parameters.
05. Establish PCR protocols using meat and bone meal-extracted and animal feed-extracted DNA.
06. Adapt PCR to the Taqman System.
07. Determine sensitivity of PCR/Taqman test.
08. Produce interim report summarising protocols to be used in coded study.
09. Carry out coded study using pre-selected samples of known species identity.
10. Preparation of final report and paper.
Time-Scale and Cost
From: 1999

To: 2001

Cost: £259,790
Contractor / Funded Organisations
Veterinary Laboratories Agency
Animal Health              
Plants and Animals              
Fields of Study
Animal Health