Comparison of DNA extraction methods for pathogenic Yersinia enterocolitica detection from meat food by nested PCR

Abstract

The objective of this work was to compare three different methods of DNA extraction from meat food, and to determine whether these methods removed inhibitors of nested PCR for pathogenic Yersinia enterocolitica detection. The amplification of the yadA gene from the DNA obtained from a pure Y. enterocolitica culture could be carried out with all the protocols. DNA amplification from the food samples was observed with two of the three tested protocols, which gave highly sensitive amplifications (detection limit 1 CFU/ml). These protocols detected a lower limit of 0.6 fg/μl of DNA extracted from Y. enterocolitica pure culture. We concluded that these protocols were able to eliminate satisfactorily the PCR inhibitors present in the foods. The nested PCR tested could be used satisfactorily in the investigation of pathogenic Y. enterocolitica in foods in the presence of a high background of microflora.