Laboratory of Plant Transgenic Technology and Biosensor, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
Biosensors (Basel). 2017 Nov 28;7(4):56. doi: 10.3390/bios7040056.
Monitoring food safety is essential for protecting the health and safety of consumers. Conventional methods used are time consuming and laborious, requiring anywhere from three to seven days to obtain results. Thus, better monitoring methods are required. In this study, a laminated lab-on-paper chip was developed, and its use for the screening of ready-to-eat seafood was demonstrated. The assay on a chip was based on loop-mediated isothermal DNA amplification (LAMP) of the gene of and fluorescence signal detection via SYBR Gold. Overall assay processes were completed in 4.5 h., (including 3.5 h. incubation for the bacteria enrichment, direct DNA amplification with no DNA extraction, and signal detection), without relying on standard laboratory facilities. Only positive samples induced fluorescence signals on chip upon illumination with UV light (λ = 460). The method has a limit of detection of 100 copies of DNA per 50 g of sample. No cross-reactivity was observed in samples contaminated with other bacteria. On-site monitoring of the seafood products using this chip revealed that one of 30 products from low sanitation vendors (3.33%) were contaminated, and these agreed with the results of PCR. The results demonstrated a benefit of this chip assay for practical on-site monitoring.
监测食品安全对于保护消费者的健康和安全至关重要。传统的方法耗时耗力,需要三到七天才能得到结果。因此,需要更好的监测方法。在本研究中,开发了一种层叠式纸上实验室芯片,并展示了其在即食海鲜筛查中的应用。芯片上的检测基于环介导等温 DNA 扩增(LAMP)扩增和通过 SYBR Gold 进行荧光信号检测。总体检测过程在 4.5 小时内完成,(包括 3.5 小时的细菌富集孵育、无需 DNA 提取的直接 DNA 扩增和信号检测),无需依赖标准实验室设备。只有阳性样本在紫外光(λ=460)照射下会在芯片上诱导荧光信号。该方法的检测限为每 50 克样品中 100 个 DNA 拷贝。在受其他细菌污染的样本中未观察到交叉反应。使用该芯片对海鲜产品进行现场监测显示,来自低卫生供应商的 30 种产品中有 1 种(3.33%)受到污染,这些结果与 PCR 结果一致。结果表明,这种芯片检测方法在实际现场监测中具有优势。