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

• Published in: Food research international 2007-06, Vol.40 (5), p.637-642
• Main Authors: Lucero Estrada, Cecilia S.M., Carmen Velázquez, Lidia del, Genaro, Silvia Di, Stefanini de Guzmán, Ana María
• Format: Article
• Language: English
• Subjects: bacterial contamination; Biological and medical sciences; Comparison; DNA; DNA extraction protocols; food contamination; Food engineering; Food industries; food pathogens; Fundamental and applied biological sciences. Psychology; General aspects; genes; Meat food; meat products; Methods of analysis, processing and quality control, regulation, standards; microbial detection; Nested PCR; phenotype; polymerase chain reaction; yadA gene; Yersinia enterocolitica
• ISSN: 0963-9969; 1873-7145
• DOI: 10.1016/j.foodres.2006.11.008

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.