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投稿时间：2023-12-20
投稿时间：2023-12-20
中文摘要: 金黄色葡萄球菌（Staphylococcus aureus）广泛分布于自然界，是导致食源性疾病的主要病原体之一。为构建一种简便且灵敏的检测金黄色葡萄球菌的方法，该文将80α 噬菌体内溶素模块中有着特异性宿主识别和结合作用的催化结构域与细胞结合域模块（Amidase 3-CBD）利用SpyCatcher/SpyTag 蛋白交联系统有方向性的固定在Ni 磁珠表面从而构建免疫磁珠Ni-SpyCatcher-SpyTag-Amidase 3-CBD 用于捕获金葡金黄色葡萄球菌，再利用等温扩增（recombinase polymerase amplification，RPA）和CRISPR/Cas12a 显色切割即可在37 ℃下2 h 内完成检测，并且人工污染牛奶样品中金黄色葡萄球菌的最低检测限为1×102 CFU/mL。该技术不需要专业技术人员或辅助设备，便可实现对金黄色葡萄球菌的高效检测。
Abstract:Staphylococcus aureus ubiquitous in the nature is one of the main pathogens causing foodborne diseases.A simple and sensitive method for detecting S.aureus was established with the catalytic domain and cellbinding domain module（Amidase 3-CBD）of the bacteriophage endolysin 80α（with the functions of specific host recognition and binding），the SpyCatcher/SpyTag system，and Ni magnetic beads. In this way，Ni-Spy-Catcher-SpyTag-Amidase 3-CBD was established for capturing S.aureus.S.aureus was then detected by recombinase polymerase amplification（RPA）and CRISPR/Cas12a chromogenic cleavage at 37 ℃.The whole detection process was completed within 2 h，and the lowest limit of detection for S. aureus in artificially contaminated milk samples was 1×102 CFU/mL. The technology facilitates efficient detection of S. aureus，dispensing with the requirement for specialized technical personnel or auxiliary equipment.
keywords: Staphylococcus aureus cell-binding domain immunomagnetic separation technology Spy-Catcher/SpyTag system recombinase polymerase amplification（RPA） CRISPR/Cas12a
文章编号：202419023 中图分类号： 文献标志码：
基金项目:广东省重点建设学科科研能力提升项目（2021ZDJS005）；广东省研究生教育创新计划项目（2023SFKC_057）
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