Di Maro Orlandina, Proroga Yolande T R, Castellano Silvia, Balestrieri Anna, Capuano Federico, Arletti Enrico, Vietina Michelangelo, Bizzarri Melissa, Murru Nicoletta, Peruzy Maria Francesca, Cristiano Daniela
Department of Food Inspection, Experimental Zooprophylactic Institute of Southern Italy, Portici.
Generon S.p.A., San Prospero.
Ital J Food Saf. 2024 Feb 26;13(1):11635. doi: 10.4081/ijfs.2024.11635. eCollection 2024 Feb 22.
The majority of human diseases attributed to seafood are caused by spp., and the most commonly reported species are , , and . The conventional methods for the detection of species involve the use of selective media, which are inexpensive and simple but time-consuming. The present work aimed to develop a rapid method based on the use of multiplex real-time polymerase chain reaction (PCR) to detect , , and in bivalve mollusks. 30 aliquots of bivalve mollusks () were experimentally inoculated with two levels of , , and . ISO 21872-1:2017 was used in parallel for qualitative analysis. The limit of detection of 50% was 7.67 CFU/g for , 0.024 CFU/g for , and 1.36 CFU/g for . For and , the real-time PCR protocol was demonstrated to amplify the pathogens in samples seeded with the lowest and highest levels. The molecular method evaluated showed a concordance rate of 100% with the reference microbiological method. was never detected in samples contaminated with the lowest level, and it was detected in 14 samples (93.33%) seeded with the highest concentration. In conclusion, the developed multiplex real-time PCR proved to be reliable for and Results for are promising, but further analysis is needed. The proposed method could represent a quick monitoring tool and, if used, would allow the implementation of food safety.
大多数归因于海鲜的人类疾病是由 物种引起的,最常报告的物种是 、 和 。检测 物种的传统方法涉及使用选择性培养基,这些方法成本低且操作简单,但耗时较长。本研究旨在开发一种基于多重实时聚合酶链反应(PCR)的快速方法,用于检测双壳贝类中的 、 和 。对30份双壳贝类( )等分试样进行了两种水平的 、 和 的实验接种。同时使用ISO 21872-1:2017进行定性分析。50%的检测限对于 为7.67 CFU/g,对于 为0.024 CFU/g,对于 为1.36 CFU/g。对于 和 ,实时PCR方案被证明能够扩增接种了最低和最高水平病原体的样本中的病原体。所评估的分子方法与参考微生物学方法的一致性率为100%。在接种最低水平污染物的样本中从未检测到 ,而在接种最高浓度的14个样本(93.33%)中检测到了 。总之,所开发的多重实时PCR被证明对 和 是可靠的。对于 的结果很有前景,但需要进一步分析。所提出的方法可以代表一种快速监测工具,如果使用,将有助于实施食品安全。