Cecere Paola, Gatto Francesca, Cortimiglia Claudia, Bassi Daniela, Lucchini Franco, Cocconcelli Pier Sandro, Pompa Pier Paolo
Nanobiointeractions & Nanodiagnostics, Istituto Italiano di Tecnologia (IIT), 16163 Genova, Italy.
Dipartimento di Scienze e Tecnologie Alimentari per la Sostenibilità della Filiera Agro-Alimentare, Facoltà di Scienze Agrarie Alimentari ed Ambientali, Università Cattolica del Sacro Cuore, Via Emilia Parmense 84, 29122 Piacenza-Cremona, Italy.
Biosensors (Basel). 2021 Aug 24;11(9):293. doi: 10.3390/bios11090293.
represents the main spoiling agent responsible for late blowing defects (LBD) in hard and semi-hard cheeses. Its spores are resistant to manufacturing procedures and can germinate during the long ripening process, causing the burst of the cheese paste with a consequent undesirable taste. The lower quality of blown cheeses leads to considerable financial losses for the producers. The early identification of spore contaminations in raw milk samples thus assumes a pivotal role in industrial quality control. Herein, we developed a point of care (POC) testing method for the sensitive detection of in milk samples, combining fast DNA extraction (with no purification steps) with a robust colorimetric loop-mediated isothermal amplification (LAMP) technique. Our approach allows for the sensitive and specific detection of spores (limit of detection, LoD: ~2 spores/mL), with the advantage of a clear naked-eye visualization of the results and a potential semi-quantitative discrimination of the contamination level. In addition, we demonstrated the feasibility of this strategy using a portable battery-operated device that allowed both DNA extraction and amplification steps, proving its potential for on-site quality control applications without the requirement of sophisticated instrumentation and trained personnel.
是导致硬质和半硬质奶酪后期产气缺陷（LBD）的主要 spoiling 因子。其孢子对制造工艺具有抗性，并且能够在漫长的成熟过程中萌发，导致奶酪糊破裂，进而产生不良味道。产气奶酪质量较低会给生产商造成相当大的经济损失。因此，原奶样品中孢子污染的早期识别在工业质量控制中起着关键作用。在此，我们开发了一种用于灵敏检测牛奶样品中 的即时检测（POC）方法，该方法将快速 DNA 提取（无需纯化步骤）与强大的比色环介导等温扩增（LAMP）技术相结合。我们的方法能够灵敏且特异性地检测 孢子（检测限，LoD：约 2 个孢子/毫升），具有结果可通过肉眼清晰可视化以及对污染水平进行潜在半定量区分的优势。此外，我们使用便携式电池供电设备证明了该策略的可行性，该设备可同时进行 DNA 提取和扩增步骤，证明了其在无需复杂仪器和专业人员的情况下用于现场质量控制应用的潜力。
