Development of a molecular methodology for yersinia enterocolitica detection in food

LAPADULA WJ; LUCERA ESTRADA C; JURI AYUB M; ECUDERO ME; MASTRODONATO AC; FAVIER G

Parana

Congreso; LIV Reunión anual sociedad Argentina de investigación en Bioquímica y biología molecular; 2018

The rapid detection of pathogenic microorganisms in foods is an essential part of the quality control to ensure the consumer health. Yersinia enterocolitica (Ye) is anenteropathogen that causes enterocolitis and extraintestinal symptoms. Its transmission is by oral route, commonly through contaminated foods. This species includes 6 biotypes, and its pathogenicity is attributed to plasmid and chromosomal virulence factors. The objective of this work was to develop a sensitive and specific methodology to detect all Ye biotypes by a one-step molecular technique. For this purpose, a conventional PCR targeted to the yst chromosomal gene, which encodes a thermostable enterotoxin associated with diarrhea in clinical cases of yersiniosis, was designed. Previous results in our laboratory demonstrated that the Ye detection limit (DL) in culture corresponded to 45 cfu/ml, while the DL obtained by PCR targeted to yst gene corresponded to 6 cfu/ml. Moreover, the results obtained in the specificity tests revealed that 19 strains of different Ye biotypes were yst , while 10 strains of other Yersinia species and 14 strains belonging to other enterobacteria were yst . Our findings indicate that the PCR based on the detection of the Yeystgene is a sensitive and specific technique which allows accelerate the detection times in comparison with the culture techniques, and might be used for the rapid detect ion of this microorganism in foods. It would be a very valuable tool to be included in a systematic surveillance of Ye in foods aimed to monitor the spread of this enteropathogen and to prevent the risk of infection in humans