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

CECILIA S. M. LUCERO ESTRADA; LIDIA DEL CARMEN VELÁZQUEZ; SILVIA DI GENARO; ANA MARÍA STEFANINI DE GUZMÁN

FOOD RESEARCH INTERNATIONAL

ELSEVIER SCIENCE BV

Año: 2007 vol. 40 p. 637 - 642

0963-9969

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 1CFU/ml). These protocols detected a lower limit of 0.6 fg/ìl 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.