Lateral flow assay of methicillin-resistant Staphylococcus aureus using bacteriophage cellular wall-binding domain as recognition agent
Lateral flow assay of methicillin-resistant Staphylococcus aureus using bacteriophage cellular wall-binding domain as recognition agent
复制标题
使用噬菌体细胞壁结合域作为识别剂对耐甲氧西林金黄色葡萄球菌进行侧流分析
DOI：
10.1016/j.bios.2021.113189
复制
发表时间：
2021-03-31
影响因子：
12.6
通讯作者：
Fu, Zhifeng
中科院分区：
文献类型：
--
作者：
Yang, Honglin;Wang, Yingran;Fu, Zhifeng
耐甲氧西林金黄色葡萄球菌（MRSA）作为最常见和最引人注目的超级细菌之一，长期以来一直是公共卫生的主要威胁。为满足对MRSA感染的有效诊断需求，迫切需要建立快速检测方法。本研究利用重组表达技术从噬菌体P108中获得了一种水溶性的细胞壁结合结构域（CWBD）蛋白。它可以作为所有MRSA菌株的广谱结合剂，并排除甲氧西林敏感的金黄色葡萄球菌菌株和其他细菌的干扰。为了建立MRSA的侧流检测（LFA）方法，以CWBD偶联的时间分辨荧光微球（FM）作为示踪MRSA的信号探针，用猪IgG固定的硝酸纤维素膜捕获MRSA。与LFA的基础上的三明治格式，MRSA可以测定在10分钟内与广泛的线性范围6.6？一百零二？6.6 ? 107 CFU/mL。它的应用潜力已被证明通过测定不同类型的细菌污染的真实的样品。结果表明，使用重组CWBD作为识别剂的LFA条提供了一种快速、便携、成本效益高的方法用于MRSA的床旁检测。
As one of the most common and noticeable superbugs, methicillin-resistant Staphylococcus aureus (MRSA) has long been a major threat to public health. To meet the demand for effective diagnosis of MRSA-induced infection, it is urgent to establish rapid assay method for this type of pathogen. In this study, an aqueous soluble cellular wall-binding domain (CWBD) protein from bacteriophage P108 was obtained with a recombinant expression technique. It can act as a wide-spectrum binding agent for all MRSA strains and exclude the interference from methicillin-susceptible strains of Staphylococcus aureus and other species of bacteria. To establish a lateral flow assay (LFA) method for MRSA, CWBD-coupled time-resolved fluorescent microspheres (FMs) were used as signal probes for tracing MRSA, and a nitrocellulose membrane immobilized with porcine IgG was used to capture MRSA. With the LFA based on sandwich format, MRSA can be assayed within 10 min with a broad linear range of 6.6 ? 102?6.6 ? 107 CFU/mL. Its application potential has been demonstrated by assaying different types of bacteria-contaminated real samples. The results suggest that the LFA strip using recombinant CWBD as the recognition agent provides a rapid, portable, cost-effective approach for point-of-care testing of MRSA.