Intralaboratory validation of meatspecies identification method was performed according to european standards and regulations. Validation was performed by designed test-system "Meat-test" on the basis of TagMan technology Real-Time PCR.This test-system allows to perform species-specific identification of animal tissues in meat raw material, grinded up meat raw material in the content of semi-finished meat products, finished untreated meat products, and meat products under thermal treatment. Test-system id designed as multiplex and it allows to identify three targets-chicken, pork and beef, simultaneously. Method validation was performed for such basic parameters as specificity, limit of detection, analytical sensitivity, amplification efficiency and linearity. Method specificity was 100% for all three indicators – Fam (chicken),Hex (pork), and Rox (beef). No cross reactions was detected. No falsenegative and false-positive results was detected, what shows high level of applied method. Limit of detection and analytical sensitivity for Fam and Hex was 1×10-6 and 1×10-5 ng, respectively. Limit of detection was 1×10-7 ng, and analytical sensitivity was 1×10-6 ng for Rox. Obtained values of slope of calibration curves (α) were within -3.58 to -3.10,what shows normal amplification efficiency. Calculated PCR efficiency values for Fam, Hex, Rox, was 103.2, 100.0 and 94.9%, respectively, what is high enough value for diagnostic test-system. Clear dependence of analysed samples Ct values on logarithm of initial concentration of added substrate was shown (R2≥0.98). Obtained validation parameters meet the EU PCR-performing standards. It enables application of this method in laboratory practice. This approach can be recommended to test laboratories for high reliability implementation of obtained results and analytical errors (false-positive and false-negative results) elimination
validation, Real-Time PCR, meat falsification
Retrieved from No. 9-10(71), 2018
Pages 96-103