Validation of a Loop-Mediated Amplification/ISO 6579-Based Method for Analysing Soya Meal for the Presence of Salmonella enterica
An alternative method for detection of Salmonella spp. in animal feed, based on the use of loop-mediated amplification (LAMP) in conjunction with a standard culturing procedure, was compared with the standard ISO 6579 as reference method, using soya meal as the test matrix. In the method comparison study, the sensitivities for both the alternative and reference methods were 100 %. The relative level of detection was 1.000. Tested against 100 Salmonella and 30 non-Salmonella strains, the LAMP-based method was 99 % inclusive and 100 % exclusive. The interlaboratory trial involved ten laboratories from eight European countries, testing eight samples at three contamination levels: 0 cfu/100 g, 1–5 cfu/100 g and 14–68 cfu/100 g. The trial specificity, or percentage correct identification of uncontaminated samples, was 96.3 % for both the reference methods and the LAMP/ISO 6579 alternative method, thus demonstrating its suitability for adoption as a procedure for rapid identification of Salmonella uncontaminated soya meal.
KeywordsSalmonella Soya meal Lamp Validation Alternative method
We thank the Q-SAFFE project partners, particularly John Thompson and Sons Ltd., for the supply of feed samples. We thank Dr. Dario de Medici of ISS for assistance with the study.
Compliance with Ethical Standards
The research leading to these results has received funding from the European Commission 7th Framework Project; Quality and Safety of Feeds and Food for Europe, QSAFFE (FP7-KBBE-2010-4, 265,702).
Conflict of Interest
Avelino Alvarez-Ordonez has no conflict of interest.
Nigel Cook has no conflict of interest.
Martin D’Agostino has no conflict of interest.
Elisabetta Delibato has no conflict of interest.
Stephen Forsythe has no conflict of interest.
Patricia Gonzalez-Garcia has no conflict of interest.
Flemming Hansen has no conflict of interest.
John Ikonomopoulos has no conflict of interest.
Emily E. Jackson has no conflict of interest.
Kieran Jordan has no conflict of interest.
Petros Kokkinos has no conflict of interest.
Krzysztof Kwiatek has no conflict of interest.
Elżbieta Kukier has no conflict of interests.
Dubravka Milanov has no conflict of interest.
Vasileios Ntafis has no conflict of interest.
Lorna O’Brien has no conflict of interest.
Tamas Petrovic has no conflict of interest.
Susana Robles has no conflict of interest.
David Rodriguez Lazaro has no conflict of interest.
Zbigniew Sieradzki has no conflict of interest.
This article does not contain any studies with human or animal subjects.
- Anonymous (2002) ISO 6579:2002—microbiology of food and animal feeding stuffs—horizontal method for the detection of Salmonella spp. International Organisation for Standardization, GenevaGoogle Scholar
- Anonymous (2016) ISO 16140–1/2—microbiology of the food chain—method validation. International Organisation for Standardization, GenevaGoogle Scholar
- Delibato E, Rodriguez-Lazaro D, Gianfranceschi M, De Cesare A, Comin D, Gattuso A, Hernandez M, Sonnessa M, Pasquali F, Sreter-Lancz Z, Saiz-Abajo MJ, Pérez-De-Juan J, Butrón J, Prukner-Radovcic E, Horvatek Tomic D, Johannessen GS, Jakočiūnė D, Olsen JE, Chemaly M, Le Gall F, González-García P, Lettini AA, Lukac M, Quesne S, Zampieron C, De Santis P, Lovari S, Bertasi B, Pavoni E, Proroga YT, Capuano F, Manfreda G, De Medici D (2014) European validation of real-time PCR method for detection of Salmonella spp. in pork meat. Int J Food Microbiol 184:134–138CrossRefGoogle Scholar
- Maurischat S, Baumann B, Martin A, Mealorny B (2014) Rapid detection and specific differentiation of Salmonella enterica subsp. enterica Enteritidis, Typhimurium and its monophasic variant 4,,12:i:—by real-time multiplex PCR. Int J Food Microbiol 79:175–181Google Scholar