School of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China.
School of Public Health, Ningbo Univesity, Ningbo 315211, China.
Anal Chem. 2024 Oct 8;96(40):16007-16016. doi: 10.1021/acs.analchem.4c03465. Epub 2024 Sep 27.
The proliferation speed of live foodborne pathogens is fast. A small number of pathogens will have a great impact on food and the environment if positive samples are not detected timely. In this study, transparent porous hydrogel stir bars, modified by two different phages (corresponding to two different bacteria ( and )), have been developed for rapid detection of foodborne bacteria. A large number of samples can be analyzed simultaneously with a small animal live imager device to screen out the positive samples, while an adenosine triphosphate (ATP) bioluminescence sensor can be used to quantify the number of bacteria in the positive samples. The phage has good specificity and capture ability to bacteria, which makes the method highly sensitive. In addition, the use of multiple phages also enables the method to detect multiple bacteria simultaneously. The three-dimensional structure of the hydrogel allows it to modify more phages, and its transparent nature also allows the inside bioluminescence to be detected. Both can enhance the sensitivity of the detection. Finally, the reagents needed for bioluminescence, such as d-luciferin, can also be preencapsulated in the hydrogel, thus simplifying the detection step. Under the best conditions, the detection range of the method is 10-10 CFU·mL, and the limit of detection is 30 CFU·mL within 11 min. The test results of actual samples show that there is no difference between using the method developed through this study and the traditional plate counting method, but the detection time is greatly shortened.
食源性致病菌的增殖速度很快。如果不能及时检测出阳性样本，少量的致病菌就会对食品和环境产生重大影响。在本研究中，开发了一种由两种不同噬菌体（分别对应两种不同细菌（和））修饰的透明多孔水凝胶搅拌棒，用于快速检测食源性细菌。可以使用小动物活体成像设备同时分析大量样本，筛选出阳性样本，同时可以使用三磷酸腺苷（ATP）生物发光传感器定量阳性样本中的细菌数量。噬菌体对细菌具有良好的特异性和捕获能力，使得该方法具有很高的灵敏度。此外，使用多种噬菌体还可以实现同时检测多种细菌。水凝胶的三维结构使其能够修饰更多的噬菌体，而其透明的性质也允许内部生物发光被检测到。这两者都可以增强检测的灵敏度。最后，生物发光所需的试剂，如 d-荧光素，也可以预先封装在水凝胶中，从而简化检测步骤。在最佳条件下，该方法的检测范围为 10-10 CFU·mL，在 11 分钟内检测限为 30 CFU·mL。实际样品的测试结果表明，使用本研究开发的方法与传统平板计数法之间没有差异，但检测时间大大缩短。
