Huang Xueqin, Chen Lingzhi, Zhi Weixia, Zeng Runmin, Ji Guanxu, Cai Huaihong, Xu Jun, Wang Jinyong, Chen Shanze, Tang Yong, Zhang Jianglin, Zhou Haibo, Sun Pinghua
The Second Clinical Medical College (Shenzhen People's Hospital), Jinan University, Shenzhen 518020, China.
The Fifth Affiliated Hospital of Jinan University, Heyuan 517000, China.
Anal Chem. 2023 Sep 5;95(35):13101-13112. doi: 10.1021/acs.analchem.3c01631. Epub 2023 Aug 1.
A new lateral flow immunoassay strip (LFIA) combining sensitive detection and identification of multiple bacteria remains a huge challenge. In this study, we first developed multifunctional urchin-shaped Au-Ag@Pt nanoparticles (UAA@P NPs) with a unique combination of colorimetric-SERS-photothermal-catalytic (CM/SERS/PT/CL) properties and integrated them with LFIA for multiplexed detection and specific discrimination of pathogenic bacteria in blood samples. Unlike the conventional LFIA that relied on antibody (Ab), this novel LFIA introduced 4-mercaptophenylboronic acid (4-MPBA) as an ideal Ab replacer that was functionalized on UAA@P NPs (UAA@P/M NPs) with outstanding binding and enrichment capacities toward bacteria. Taking () as model bacteria, the limit of detection (LOD) was 3 CFU/mL for SERS-LFIA, 27 CFU/mL for PT-LFIA, and 18 CFU/mL for CL-LFIA, three of which were over 330-fold, 37-fold, and 55-fold more sensitive than ordinary visual CM-LFIA, respectively. Besides, this SERS-LFIA is capable of identifying three types of bacterial spiked blood samples (, , and ) effectively according to specific bacterial Raman "fingerprints" by partial least-squares-discriminant analysis (PLS-DA). More importantly, this LFIA was successfully applied to blood samples with satisfactory recoveries from 90.3% to 108.8% and capable of identifying the infected patients ( = 4) from healthy subjects ( = 2) with great accuracy. Overall, the multimodal LFIA incorporates bacteria discrimination and quantitative detection, offering an avenue for early warning and diagnosis of bacterial infection.
开发一种能够同时灵敏检测和鉴定多种细菌的新型侧向流动免疫分析试纸条(LFIA)仍然是一项巨大的挑战。在本研究中,我们首先制备了具有比色-表面增强拉曼光谱-光热-催化(CM/SERS/PT/CL)独特组合性能的多功能海胆状金-银@铂纳米颗粒(UAA@P NPs),并将其与LFIA集成,用于血液样本中病原菌的多重检测和特异性鉴别。与传统的依赖抗体(Ab)的LFIA不同,这种新型LFIA引入了4-巯基苯硼酸(4-MPBA)作为理想的抗体替代物,其在UAA@P NPs(UAA@P/M NPs)上功能化,对细菌具有出色的结合和富集能力。以()作为模式细菌,SERS-LFIA的检测限(LOD)为3 CFU/mL,PT-LFIA为27 CFU/mL,CL-LFIA为18 CFU/mL,这三种检测方法的灵敏度分别比普通视觉CM-LFIA高330倍、37倍和55倍以上。此外,这种SERS-LFIA能够通过偏最小二乘判别分析(PLS-DA),根据特定细菌的拉曼“指纹”有效识别三种添加了细菌的血液样本(,和)。更重要的是,这种LFIA成功应用于血液样本,回收率在90.3%至108.8%之间,能够准确地从健康受试者(=2)中识别出感染患者(=4)。总体而言,这种多模式LFIA结合了细菌鉴别和定量检测,为细菌感染的早期预警和诊断提供了一条途径。