Zhang Guangying, Zhang Lili, Ye Jingqin, Wang Dongshu, Lu Ying
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai 201306, China.
Biosensors (Basel). 2025 Aug 21;15(8):553. doi: 10.3390/bios15080553.
is the main spoilage bacterium in milk, and its proliferation is one of the factors leading to the deterioration of the quality of raw milk. In this study, a rapid detection system for was developed based on recombinase-aided amplification combined with a test strip (RAA-TS), which contained a double test line (DTL) targeting the virulence gene of and the housekeeping gene of . Visual observation could detect (50 CFU/mL) and (250 CFU/mL) within 90 min, including sample pretreatment and RAA reaction and detection steps. No cross-reactions were observed with or other bacteria (n = 19). The quantitative detection limits (LOD) of and for in milk were 37 CFU/mL and 233 CFU/mL, respectively. Compared with polymerase chain reaction-agarose gel electrophoresis (PCR-AGE), the sensitivity of the developed RAA-TS-DTL system was increased by approximately four times. Furthermore, it could detect live in milk when combined with optimized sample pretreatment by propidium monoazide (PMAxx). Its consistency with the traditional culture method in the detection of spiked in milk samples (n = 25) was 100%. The developed RAA-TS-DTL had the advantages of high accuracy and short time consumption. Thus, it provides a new way or tool for the rapid screening or detection of in milk.
是牛奶中的主要腐败细菌，其增殖是导致原料奶质量下降的因素之一。在本研究中，基于重组酶辅助扩增结合试纸条（RAA-TS）开发了一种用于的快速检测系统，该试纸条包含针对的毒力基因和的管家基因的双检测线（DTL）。通过目视观察可在90分钟内检测到（50 CFU/mL）和（250 CFU/mL），包括样品预处理以及RAA反应和检测步骤。与或其他细菌（n = 19）未观察到交叉反应。牛奶中对和的定量检测限（LOD）分别为37 CFU/mL和233 CFU/mL。与聚合酶链反应-琼脂糖凝胶电泳（PCR-AGE）相比，所开发的RAA-TS-DTL系统的灵敏度提高了约四倍。此外，当与单叠氮化丙锭（PMAxx）优化的样品预处理相结合时，它可以检测牛奶中的活。其在检测牛奶样品（n = 25）中加标的与传统培养方法的一致性为100%。所开发的RAA-TS-DTL具有准确性高和耗时短的优点。因此，它为牛奶中的快速筛选或检测提供了一种新方法或工具。
