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

被引：24
作者：
Cammilleri, Gaetano [1 ,2 ]
Ferrantelli, Vincenzo [1 ]
Pulvirenti, Andrea [2 ]
Drago, Chiara [3 ]
Stampone, Giuseppe [3 ]
Quintero Macias, Gema Del Rocio [3 ]
Drago, Sandro [3 ]
Arcoleo, Giuseppe [3 ]
Costa, Antonella [1 ]
Geraci, Francesco [1 ]
Di Bella, Calogero [1 ]
机构：
[1] Ist Zooprofilatt Sperimentale Sicilia, Via Gino Marinuzzi 3, I-90129 Palermo, Italy
[2] Univ Modena & Reggio Emilia, Dipartimento Sci Vita, Via Univ 4, I-41121 Modena, Italy
[3] Enbiotech Srl, Aquileia 34, I-90144 Palermo, Italy
来源：
FOODS | 2020年 / 9卷 / 01期
关键词：
Anisakis spp; molecular methods; LAMP; validation; anisakidae family; REAL-TIME PCR; PARASITE ANISAKIS; SANDWICH ELISA; SIMPLEX; NEMATODES; PEGREFFII; LARVAE; QUANTIFICATION; DIAGNOSIS; ALLERGENS;
D O I：
10.3390/foods9010092
中图分类号：
TS2 [食品工业];
学科分类号：
0832 ;
摘要：
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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页数：11
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