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Development of methods for the identification of duck, pheasant, venison, horse, donkey and wild boar in meat products (Q01083) - FA0208

Description
The objective of this study was to design and optimise real-time PCR assays for the specific detection of horse, donkey, duck, deer, pheasant and wild boar in commercial products.

The assays were to be easy to use and applicable for use by public analysts and other laboratories. Although the assays were primarily for use with real-time PCR equipment, they were also to be available for use by laboratories which did not have real-time PCR capability, but which did have experience of gel electrophoresis. Finally, the outputs of the project were to include a comprehensive set of standard operating procedures to facilitate technology transfer to other laboratories.

The objectives of the study were broadly met, with assays specific for the detection of all target species successfully designed and optimised, with the exception of wild boar. After extensive sequence analysis, and utilising the results of population genetics work at CSL (a MAFF funded project), it was concluded that the identification of wild boar in commercial products was not achievable using this real-time PCR based approach and that a microsatellite based approach might be more applicable.

The real-time PCR based approach was found to be applicable to the detection of duck, deer, horse, donkey and pheasant. Sensitive and specific assays were designed and optimised which were shown to be capable of detecting their target species in heat and pressure processed commercial products. The design of the assays made them fully portable between real-time PCR and gel electrophoresis equipment. Finally technology transfer has been facilitated by the early submission of two manuscripts to peer reviewed journals detailing the design and optimisation of the horse, donkey and duck assays, both of which have been accepted for publication. Additionally, a comprehensive set of standard operating procedures has been provided for use by public analysts and other laboratories.

The final part of this project sought to evaluate an emerging methodology for species identification which utilises chip technology. The Food Expert ID system, developed by bioMerieux, was loaned to CSL for a period of 6 months and its operation and applicability to the identification of species in food products evaluated. The range of species included on the chip allowed samples from 3 FSA funded projects to be assayed: white fish mixtures from the CCFRA fish project, meat mixtures from the DNA quality project (Q01033 & Q01034) and exotic meats from the Exotic meat project (Q01083). Additionally the chip was assessed using tuna standards, originally from the FSA tuna survey, conducted in 1999. The system was able to detect both single species and species in mixtures, even after heat and pressure processing. Cod was detected in a mixture of cod and haddock, and pork, beef and chicken were also detected, all below the 5% evel given as the sensitivity of the assay. A range of tuna species were correctly identified from raw muscle, however canned tuna could not be detected, even at 100%. The system was easy to use and provided information on the species present in a samples without running numerous assays. When the technology becomes more established it is anticipated that the range of species included on the chip will be extended and the price of the chips fall, making these types of analysis more widely available.
Objective
1. Objective –Acquisition of mCyt b sequence data for duck, pheasant, wild boar, venison, donkey and horse
2. Objective – Design and optimisation of primers and probes for duck, pheasant, wild boar, venison, donkey and horse
3. Objective –Determine the specificity and limit of detection of each primer set
4. Objective –Validate assays
5. Objective – To determine the performance characteristics of the BioMerieux food Expert ID system.
6. Objective - Technology transfer

Project Documents
• EVID4 - Final project report : Q01083 RCUA5   (198k)
• FRP - Final Report : final report Q01083   (473k)
• OTH - Other : FSA SOP Common Exotic Meats (Tepnel ext) August 2009 V4   (225k)
• OTH - Other : FSA SOP common exotic meat CTAB extraction   (265k)
• OTH - Other : FSA SOP common exotic meat tepnel extraction   (208k)
• OTH - Other : FSA SOP common exotic meat tepnel extraction   (208k)
Time-Scale and Cost
From: 2003

To: 2005

Cost: £31,358
Contractor / Funded Organisations
F E R A (FERA)
Keywords
Food