Wang Liping, Wu Ziyun
School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200030, China.
Foods. 2024 Sep 18;13(18):2954. doi: 10.3390/foods13182954.
Normal and damaged microorganisms are related to food safety. The colony-forming unit (CFU) assay and viability of microorganisms have broad applications in food. Traditionally, the CFU assay has been the benchmark for assessing microbial viability across various fields. However, the normal and damaged microorganisms cannot be distinguished. Here, we introduce an improved technology for foods that uses a visible absorbance microplate reader platform for high-throughput quantitative analysis of microbial lag time, doubling time, and CFU. This platform utilizes a 96-well plate and a microplate reader to accurately determine the viable cell number from a five-microliter sample. It boasts the capability to measure a dynamic range spanning from five to seven orders of magnitude, significantly reducing the time required by over 20-fold in comparison to traditional spread plate methods. Additionally, it demonstrates a remarkable ability to detect a single cell within a well. A mild temperature treatment for cell viability detection was implemented and was able to reflect the real microbial quality. Consequently, the high-throughput method as an improved technology provides essential technical support for microbial detection.
正常微生物和受损微生物与食品安全相关。菌落形成单位(CFU)测定法和微生物活力在食品领域有广泛应用。传统上,CFU测定法一直是评估各个领域微生物活力的基准。然而,正常微生物和受损微生物无法区分。在此,我们介绍一种针对食品的改进技术,该技术使用可见吸光酶标仪平台对微生物延迟期、倍增时间和CFU进行高通量定量分析。该平台利用96孔板和酶标仪从五微升样品中准确测定活细胞数量。它能够测量跨越五个到七个数量级的动态范围,与传统平板涂布法相比,所需时间显著减少20多倍。此外,它在检测孔内单个细胞方面表现出卓越能力。实施了用于细胞活力检测的温和温度处理,并且能够反映真实的微生物质量。因此,作为一种改进技术的高通量方法为微生物检测提供了重要的技术支持。