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Evaluation of ISO enrichment real‐time PCR methods with internal amplification control for detection of Listeria monocytogenes and Salmonella enterica in fresh fruit and vegetables
Author(s) -
Badosa E.,
Chico N.,
Pla M.,
Parés D.,
Montesinos E.
Publication year - 2009
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2009.02628.x
Subject(s) - listeria monocytogenes , salmonella enterica , salmonella , biology , detection limit , real time polymerase chain reaction , food science , listeria , polymerase chain reaction , microbiology and biotechnology , bacteria , chromatography , chemistry , gene , biochemistry , genetics
Aims: To provide with a quick method for qualitative detection, in less than three days, of Salmonella enterica and Listeria monocytogenes in fresh fruit and vegetables. Methods and Results: The method was based on coupling International Standard Organization (ISO) enrichment to a real‐time PCR with internal amplification control (IAC), in a duplex format, without additional DNA purification. The performance was tested on different plant products. Both bacterial pathogens were consistently detected with a limit of detection (LOD) of 1 CFU in 25 g after enrichment, except for soybean sprouts. Levels of S. enterica , ranging from 1 to 10 CFU in 25 g after enrichment were detected with different enrichment broths. Conclusions: For both pathogens, the LOD was similar to that of the corresponding ISO method, while decreasing the analysis time and handling needs. Significance and Impact of the Study: The agreement between standard ISO and the enrichment real‐time PCR(IAC)‐based methods make the latter method as a promising alternative for quick and reliable detection of food‐borne pathogens in fresh fruit and vegetables in routine laboratories.