Bio-hybrid nanoarchitectonics of nanoflower-based ELISA method for the detection of Staphylococcus aureus
Bio-hybrid nanoarchitectonics of nanoflower-based ELISA method for the detection of Staphylococcus aureus
复制标题
基于纳米花的生物混合纳米结构酶联免疫吸附法检测金黄色葡萄球菌
DOI：
10.1016/j.snb.2022.132005
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发表时间：
2022-05-11
影响因子：
8.4
通讯作者：
He, Jin
中科院分区：
文献类型：
--
作者：
Yin, Wen;Zhu, Li;He, Jin
金黄色葡萄球菌是引起食物中毒和人类传染病的重要病原菌之一。因此，建立一种快速、灵敏的沙门氏菌检测方法具有重要意义。金黄色葡萄球菌对诊断、预防和控制由该病原体引起的疾病具有重要意义。为此，我们建立了一种基于纳米花的生物杂化纳米结构的ELISA检测方法。金黄色葡萄球菌具有高度的特异性和敏感性。该方法的核心是通过纳米构筑制备特异性的细胞壁结合域（CBD）-辣根过氧化物酶（HRP）-Cu 3（PO 4）2三合一有机-无机杂化纳米花复合物。检测包括两个主要步骤：我们首先包被抗S。金黄色葡萄球菌多克隆抗体进入96孔板的威尔斯孔中以特异性捕获金黄色葡萄球菌。金黄色葡萄球菌，然后添加定制的CBD-HRPCu 3（PO 4）2有机-无机杂化纳米花到系统中。由于来自噬菌体phi 1的溶素PlyV 12的CBD与S.金黄色葡萄球菌，形成稳定的抗体-细菌-杂交纳米花复合物。然后，利用TMB（3，3 '，5，5'-四甲基联苯胺）-过氧化氢报告系统对杂交纳米花中HRP酶活性进行信号放大。该方法能特异性地检测S.金黄色葡萄球菌在101至106个菌落形成单位（CFU）/mL范围内线性变化，检测限低至6 CFU/mL，这可以说是迄今报道的所有检测方法中最好的方法之一。
Staphylococcus aureus is one of the crucial bacterial pathogens that cause food poisoning and human infectious diseases. Therefore, the establishment of a rapid and sensitive detection method for S. aureus is of great significance for the diagnosis, prevention, and control of the diseases caused by this pathogen. To this end, we have established a bio-hybrid nanoarchitectonics of nanoflower-based ELISA method for the detection of S. aureus with high specificity and sensitivity. The core of the method is to prepare a specific cell wall binding domain (CBD)-horseradish peroxidase (HRP)-Cu3(PO4)2 three-in-one organic-inorganic hybrid nanoflower complex through nanoarchitectonics. Detection consists of two main steps: we firstly coated an anti-S. aureus polyclonal antibody into the wells of a 96-well plate to specifically capture S. aureus, and then added customized CBD-HRPCu3(PO4)2 organic-inorganic hybrid nanoflower to the system. Due to the high specificity of the CBD from lysin PlyV12 of bacteriophage phi 1 with S. aureus, a stable antibody-bacterium-hybrid nanoflower complex was formed. Then, the signal amplification of HRP enzyme activity in the hybrid nanoflower was carried out by using the TMB (3,3 ',5,5 '-tetramethylbenzidine)-hydrogen peroxide reporting system. The method has been proved to be capable of specifically detecting S. aureus ranging from 101 to 106 conoly-forming unit (CFU)/mL linearly with a detection limit down to 6 CFU/mL, which is arguably one of the best for all detection methods reported to date.