He Dongxia, Liu Qi, Wang Lei, Han Qinqin, Zhang Jinyang, Li Chao, Song Yuzhu
Faculty of Life Science and Technology, Kunming University of Science and Technology, No.727, Jingming South Road, Chenggong District, Kunming City, 650500, China.
Anal Bioanal Chem. 2025 Apr 12. doi: 10.1007/s00216-025-05862-8.
A novel strategy integrating Brevinin-1BW (BW)-functionalized magnetic bead-based separation with a colorimetric immunoassay was developed to concentrate and enrich Staphylococcus aureus (S. aureus) from sample matrices using magnetic beads functionalized with antimicrobial peptide BW (MBs-BW) as carriers. To ensure the selectivity of the method, horseradish peroxidase (HRP)-labeled porcine IgG (HRP-porcine IgG) was used as a molecular recognition reagent and signal amplification probe. MBs-BW/S. aureus/HRP-porcine IgG probe sandwich complex was successfully obtained. HRP chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) was used to realize the chromogenic reaction. Finally, S. aureus was quantitatively analyzed based on the color signal generated. Under the optimal conditions, the assay showed a response in the concentration range of 1.0 × 10-1.0 × 10 CFU/mL of S. aureus, and the detection limit was as low as 60 CFU/mL. This method does not require complicated sample pretreatment, and the operation process is fast and simple, which can directly complete the whole process of bacterial enrichment and detection. The recovery of this strategy in different types of spiked samples reached 86.8-97.8%, and the analytical results of this strategy were highly consistent with those of the conventional plate counting method. This strategy provides a new way for the rapid detection of S. aureus in complex samples.
开发了一种将布雷维宁 - 1BW（BW）功能化磁珠分离与比色免疫分析相结合的新策略，以抗菌肽BW功能化的磁珠（MBs - BW）为载体，从样品基质中富集金黄色葡萄球菌（S. aureus）。为确保该方法的选择性，使用辣根过氧化物酶（HRP）标记的猪IgG（HRP - 猪IgG）作为分子识别试剂和信号放大探针。成功获得了MBs - BW/S. aureus/HRP - 猪IgG探针夹心复合物。使用HRP显色底物3,3',5,5'-四甲基联苯胺（TMB）实现显色反应。最后，基于产生的颜色信号对金黄色葡萄球菌进行定量分析。在最佳条件下，该分析方法对金黄色葡萄球菌的响应浓度范围为1.0×10 - 1.0×10 CFU/mL，检测限低至60 CFU/mL。该方法无需复杂的样品预处理，操作过程快速简单，可直接完成细菌富集和检测的全过程。该策略在不同类型加标样品中的回收率达到86.8 - 97.8%，其分析结果与传统平板计数法高度一致。该策略为复杂样品中金黄色葡萄球菌的快速检测提供了一种新方法。
