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基于金壳二氧化硅核纳米球的表面增强拉曼散射侧向流动免疫分析法对O157:H7的快速、定量、高灵敏度检测

Rapid, Quantitative, High-Sensitive Detection of O157:H7 by Gold-Shell Silica-Core Nanospheres-Based Surface-Enhanced Raman Scattering Lateral Flow Immunoassay.

作者信息

Shi Luoluo, Xu Ling, Xiao Rui, Zhou Zihui, Wang Chongwen, Wang Shengqi, Gu Bing

机构信息

Medical Technology Institute of Xuzhou Medical University, Xuzhou, China.

Beijing Institute of Radiation Medicine, Beijing, China.

出版信息

Front Microbiol. 2020 Nov 6;11:596005. doi: 10.3389/fmicb.2020.596005. eCollection 2020.

Abstract

O157:H7 is regarded as one of the most harmful pathogenic microorganisms related to foodborne diseases. This paper proposes a rapid-detection biosensor for the sensitive and quantitative analysis of O157:H7 in biological samples by surface-enhanced Raman scattering (SERS)-based lateral flow immunoassay (LFIA). A novel gold-shell silica-core (SiO/Au) nanosphere (NP) with monodispersity, good stability, and excellent SERS activity was utilized to prepare high-performance tags for the SERS-based LFIA system. The SiO/Au SERS tags, which were modified with two layers of Raman reporter molecules and monoclonal antibodies, effectively bind with O157:H7 and form sandwich immune complexes on the test lines. O157:H7 was quantitatively detected easily by detecting the Raman intensity of the test lines. Under optimal conditions, the limit of detection (LOD) of the SiO/Au-based SERS-LIFA strips for the target bacteria was 50 cells/mL in PBS solution, indicating these strips are 2,000 times more sensitive than colloidal Au-based LFIA strips. Moreover, the proposed assay demonstrated high applicability in O157:H7 detection in biological samples, including tap water, milk, human urine, lettuce extract and beef, with a low LOD of 100 cells/mL. Results indicate that the proposed SERS-based LFIA strip is applicable for the sensitive and quantitative determination of O157:H7.

摘要

O157:H7被认为是与食源性疾病相关的最有害致病微生物之一。本文提出了一种基于表面增强拉曼散射(SERS)的侧向流动免疫分析(LFIA)快速检测生物传感器,用于生物样品中O157:H7的灵敏定量分析。利用一种具有单分散性、良好稳定性和优异SERS活性的新型金壳二氧化硅核(SiO/Au)纳米球(NP)制备用于基于SERS的LFIA系统的高性能标签。用两层拉曼报告分子和单克隆抗体修饰的SiO/Au SERS标签与O157:H7有效结合,并在检测线上形成夹心免疫复合物。通过检测检测线的拉曼强度可轻松定量检测O157:H7。在最佳条件下,基于SiO/Au的SERS-LIFA试纸条对目标细菌在PBS溶液中的检测限为50个细胞/mL,表明这些试纸条比基于胶体金的LFIA试纸条灵敏2000倍。此外,所提出的检测方法在包括自来水、牛奶、人尿、生菜提取物和牛肉在内的生物样品中的O157:H7检测中显示出高适用性,检测限低至100个细胞/mL。结果表明,所提出的基于SERS的LFIA试纸条适用于O157:H7的灵敏定量测定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb1d/7677456/1b48996236d0/fmicb-11-596005-g006.jpg

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