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Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products

Parasites belonging to the Anisakis genera are organisms of interest for human health because they are responsible for the Anisakiasis zoonosis, caused by the ingestion of raw or undercooked fish. Furthermore, several authors have reported this parasite to be a relevant inducer of acute or chronic a...

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Autores principales: Cammilleri, Gaetano, Ferrantelli, Vincenzo, Pulvirenti, Andrea, Drago, Chiara, Stampone, Giuseppe, Del Rocio Quintero Macias, Gema, Drago, Sandro, Arcoleo, Giuseppe, Costa, Antonella, Geraci, Francesco, Di Bella, Calogero
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022600/
https://www.ncbi.nlm.nih.gov/pubmed/31963107
http://dx.doi.org/10.3390/foods9010092
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author Cammilleri, Gaetano
Ferrantelli, Vincenzo
Pulvirenti, Andrea
Drago, Chiara
Stampone, Giuseppe
Del Rocio Quintero Macias, Gema
Drago, Sandro
Arcoleo, Giuseppe
Costa, Antonella
Geraci, Francesco
Di Bella, Calogero
author_facet Cammilleri, Gaetano
Ferrantelli, Vincenzo
Pulvirenti, Andrea
Drago, Chiara
Stampone, Giuseppe
Del Rocio Quintero Macias, Gema
Drago, Sandro
Arcoleo, Giuseppe
Costa, Antonella
Geraci, Francesco
Di Bella, Calogero
author_sort Cammilleri, Gaetano
collection PubMed
description Parasites belonging to the Anisakis genera are organisms of interest for human health because they are responsible for the Anisakiasis zoonosis, caused by the ingestion of raw or undercooked fish. Furthermore, several authors have reported this parasite to be a relevant inducer of acute or chronic allergic diseases. In this work, a rapid commercial system based on Loop-Mediated Isothermal Amplification (LAMP) was optimised and validated for the sensitive and rapid detection of Anisakis spp. DNA in processed fish products. The specificity and sensitivity of the LAMP assay for processed fish samples experimentally infected with Anisakis spp. larvae and DNA were determined. The LAMP system proposed in this study was able to give positive amplification for all the processed fish samples artificially contaminated with Anisakis spp., giving sensitivity values equal to 100%. Specificity tests provided no amplification for the Contracaecum, Pseudoterranova, or Hysterothylacium genera and uninfected samples. The limit of detection (LOD) of the LAMP assay proposed was 10(2) times lower than the real-time PCR method compared. To the best of our knowledge, this is the first report regarding the application of the LAMP assay for the detection of Anisakis spp. in processed fish products. The results obtained indicate that the LAMP assay validated in this work could be a reliable, easy-to-use, and convenient tool for the rapid detection of Anisakis DNA in fish product inspection.
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spelling pubmed-70226002020-03-09 Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products Cammilleri, Gaetano Ferrantelli, Vincenzo Pulvirenti, Andrea Drago, Chiara Stampone, Giuseppe Del Rocio Quintero Macias, Gema Drago, Sandro Arcoleo, Giuseppe Costa, Antonella Geraci, Francesco Di Bella, Calogero Foods Article Parasites belonging to the Anisakis genera are organisms of interest for human health because they are responsible for the Anisakiasis zoonosis, caused by the ingestion of raw or undercooked fish. Furthermore, several authors have reported this parasite to be a relevant inducer of acute or chronic allergic diseases. In this work, a rapid commercial system based on Loop-Mediated Isothermal Amplification (LAMP) was optimised and validated for the sensitive and rapid detection of Anisakis spp. DNA in processed fish products. The specificity and sensitivity of the LAMP assay for processed fish samples experimentally infected with Anisakis spp. larvae and DNA were determined. The LAMP system proposed in this study was able to give positive amplification for all the processed fish samples artificially contaminated with Anisakis spp., giving sensitivity values equal to 100%. Specificity tests provided no amplification for the Contracaecum, Pseudoterranova, or Hysterothylacium genera and uninfected samples. The limit of detection (LOD) of the LAMP assay proposed was 10(2) times lower than the real-time PCR method compared. To the best of our knowledge, this is the first report regarding the application of the LAMP assay for the detection of Anisakis spp. in processed fish products. The results obtained indicate that the LAMP assay validated in this work could be a reliable, easy-to-use, and convenient tool for the rapid detection of Anisakis DNA in fish product inspection. MDPI 2020-01-16 /pmc/articles/PMC7022600/ /pubmed/31963107 http://dx.doi.org/10.3390/foods9010092 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cammilleri, Gaetano
Ferrantelli, Vincenzo
Pulvirenti, Andrea
Drago, Chiara
Stampone, Giuseppe
Del Rocio Quintero Macias, Gema
Drago, Sandro
Arcoleo, Giuseppe
Costa, Antonella
Geraci, Francesco
Di Bella, Calogero
Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products
title Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products
title_full Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products
title_fullStr Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products
title_full_unstemmed Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products
title_short Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products
title_sort validation of a commercial loop-mediated isothermal amplification (lamp) assay for the rapid detection of anisakis spp. dna in processed fish products
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022600/
https://www.ncbi.nlm.nih.gov/pubmed/31963107
http://dx.doi.org/10.3390/foods9010092
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