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Abstract：
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 102 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.
摘要：
属于异尖病属的寄生虫是人类健康感兴趣的生物，因为它们负责异尖病人畜共患病，由摄入生鱼或未煮熟的鱼引起的。此外，一些作者报道这种寄生虫是急性或慢性过敏性疾病的相关诱导剂。在这项工作中,优化并验证了基于环介导等温扩增（LAMP）的快速商业系统，以灵敏，快速地检测Anisakisspp。加工鱼产品中的DNA。LAMP测定法对实验感染Anisakisspp的加工鱼样品的特异性和敏感性。确定幼虫和DNA。本研究中提出的LAMP系统能够对人工污染Anisakisspp的所有加工鱼类样品进行阳性扩增。,给出的灵敏度值等于100%。特异性测试没有提供对收缩肠的扩增，伪erranova,或赤霉属和未感染的样品。建议的LAMP测定法的检测限（LOD）比比较的实时PCR方法低102倍。据我们所知,这是有关LAMP测定法在Anisakisspp检测中的应用的第一份报告。在加工鱼产品中。获得的结果表明，在这项工作中验证的LAMP测定法可能是可靠的，易于使用，和方便的工具，用于鱼产品检测中的AnisakisDNA的快速检测。
