Evaluation of a New Loop-Mediated Isothermal Amplification (LAMP) Assay for the Detection of Anisakis spp.

Tirloni, E.^1*, Stella, S.¹, Drago, C.², Stampone, G.², Vasconi, M.¹, Coppola, C.¹, Caffara, M.³, Gustinelli, A.³ & Bernardi, C.¹

¹Department of Health, Animal Science and Food Safety, Università degli Studi di Milano, Via Celoria 10, IT-20133, Milan, Italy
²Enbiotech srl, Via Aquileia, 34, IT- 90144 Palermo, Italy
³Department of Veterinary Medical Sciences, Alma Mater Studiorum University of Bologna, Via Tolara di Sopra 50, Ozzano Emilia - Italy

Dr. Erica Tirloni, Department of Health, Animal Science and Food Safety, Università degli Studi di Milano, Via Celoria 10, IT-20133, Milan, Italy.

Keywords: LAMP; Anisakis; Fish Products; Diagnostic Tool; Baby Food; Minimum Concentration Detected

Abstract

Objective of the present study was to test the performances of a real-time LAMP-based fieldfriendly tool system for the detection of Anisakis spp., with particular focus on fish products. The specificity of the method was evaluated on Anisakis spp. larvae from internal collection. 100% of the Anisakis spp. strains tested were recognized, while, correctly, no amplification occurred for non-pathogenic Hysterothylacium spp. The sensitivity was evaluated in three independent trails conducted on intentionally infested at several intensities salmon fish fillets homogenate, on seabream commercial baby food and on domestic seabream baby food. Results obtained showed a detected minimum intensity of 20 larvae/kg in the first trial (infected salmon fillets homogenate), while in intentionally infected commercial and homemade seabream baby foods this minimum intensity was 4 larvae/kg, in agreement with the limit suggested by the Codex Alimentarius for instruments intended for the identification of the presence of larvae in fishery products (5 larvae/kg). The system did not give the same performances in the equivalent matrixes after thermal treatment inactivation. This LAMP method can be considered a very useful tool for the application to fish raw matrixes as it is a cost-effective and easy-functioning method, while in the detection of inactivated larvae for the prevention of possible allergic reactions, other studies should be performed.

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