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
Istituto Zooprofilattico Sperimentale della Sicilia, via Gino Marinuzzi 3, 90129 Palermo, Italy.
Dipartimento di Scienze della Vita, Università degli studi di Modena e Reggio Emilia, Via Università 4, 41121 Modena, Italy.
Foods. 2020 Jan 16;9(1):92. doi: 10.3390/foods9010092.
Parasites belonging to the 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 spp. DNA in processed fish products. The specificity and sensitivity of the LAMP assay for processed fish samples experimentally infected with 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 spp., giving sensitivity values equal to 100%. Specificity tests provided no amplification for the , , or genera and uninfected samples. The limit of detection (LOD) of the LAMP assay proposed was 10 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 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 DNA in fish product inspection.
属于该属的寄生虫是对人类健康有重要意义的生物,因为它们会导致因食用生的或未煮熟的鱼类而引发的异尖线虫病。此外,几位作者报告称这种寄生虫是急性或慢性过敏性疾病的相关诱发因素。在这项工作中,基于环介导等温扩增(LAMP)的快速商业系统被优化并验证用于在加工鱼制品中灵敏快速地检测该属物种的DNA。测定了LAMP检测法对实验感染该属物种幼虫和DNA的加工鱼样品的特异性和灵敏度。本研究中提出的LAMP系统能够对所有人工污染该属物种的加工鱼样品进行阳性扩增,灵敏度值达100%。特异性测试对该属、该属或该属以及未感染的样品均无扩增。所提出的LAMP检测法的检测限比实时荧光定量PCR方法低10倍。据我们所知,这是关于LAMP检测法应用于检测加工鱼制品中该属物种的首次报告。所获得的结果表明,本研究中验证的LAMP检测法可能是鱼制品检测中快速检测该属DNA的一种可靠、易用且便捷的工具。