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The adaptation and validation of real-time PCR methods for the identification of exotic species, for analysis on a capillary electrophoresis chip system - FA0209

Description
The objective of this study was to adapt and validate 10 species-specific real-time
PCR assays for use on the Agilent BioAnalyser. The assays, for the detection of
horse, donkey, duck, deer, pheasant, turkey, chicken, pig, cow and sheep, had been
developed during projects previously funded by the Food Standards Agency. The
assays were to be easy to use and applicable for use by Public Analysts and other
laboratories which did not have real-time PCR capability, but which did have the
BioAnalyser. 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 fully met, with assays specific for the detection of all
target species successfully adapted and validated for use on the BioAnalyser. The
adaptation did not turn out to be as straight forward as it seemed at first, since 5 of
the 10 assays needed to be redesigned. Nevertheless, these new assays were
found to be specific and sensitive following optimisation. The assays were validated
using a variety of matrices and were found to be able to detect all species when
assessed against raw meat samples spiked at 0.5%. The assays were also found to
be capable of detecting their target species in commercial products which had been
processed, including salami and pate samples. However, the assays were found to
be relatively insensitive for the analysis of canned samples. A limited range of
canned samples (5, 10, 90 and 95% target species) where assessed and it was
found that the DNA in these samples was too degraded for the detection of target
species below 10%, although target species could be detected when present above
90%.
In conclusion, the project successfully adapted and validated assays for the detection
of 10 meat species using the Agilent BioAnalyser, providing a comprehensive
standard operating procedure to facilitate technology transfer to the Public Analysts
and other laboratories.
Project Documents
• FRP - Final Report : final report Q01107 16may08   (538k)
Time-Scale and Cost
From: 2006

To: 2007

Contractor / Funded Organisations
Keywords