Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, Iksan 54538, Korea.
College of Pharmacy, Wonkwang University, Iksan 54538, Korea.
Int J Mol Sci. 2022 Jul 19;23(14):7957. doi: 10.3390/ijms23147957.
Globally, point-of-care testing (POCT) is the most preferable on-site technique for disease detection and includes a rapid diagnostic test (RDT) and fluorescent immunochromatographic strip test (FICT). The testing kits are generally insufficient in terms of signal enhancement, which is a major drawback of this approach. Sensitive and timely on-site POCT methods with high signal enhancement are therefore essential for the accurate diagnosis of infectious diseases. Herein, we prepare cysteamine-gold coated carboxylated europium chelated nanoparticle (Cys Au-EuNPs)-mediated POCT for the detection of the H5N1 avian influenza virus (AIV). Commercial nanoparticles were used for comparison. The spectral characteristics, surface morphologies, functional groups, surface charge and stability of the Cys AuNPs, EuNPs, and Cys Au-EuNPs were confirmed by UV-visible spectrophotometry, fluorescence spectrometry, transmission electron microscope with Selected area electron diffraction (TEM-SAED), Fourier-transform infrared spectroscopy (FTIR) and zeta potential analysis. The particle size distribution revealed an average size of ~130 ± 0.66 nm for the Cys Au-EuNPs. The Cys Au-EuNP-mediated RDT (colorimetric analysis) and FICT kit revealed a limit of detection (LOD) of 10 HAU/mL and 2.5 HAU/mL, respectively, for H5N1 under different titer conditions. The obtained LOD is eight-fold that of commercial nanoparticle conjugates. The photo luminance (PL) stability of ~3% the Cys Au-EuNPs conjugates that was obtained under UV light irradiation differs considerably from that of the commercial nanoparticle conjugates. Overall, the developed Cys Au-EuNPs-mediated dual-mode POCT kit can be used as an effective nanocomposite for the development of on-site monitoring systems for infectious disease surveillance.
全球范围内,即时检测(POCT)是最受欢迎的现场检测技术,包括快速诊断检测(RDT)和荧光免疫层析条检测(FICT)。这些检测试剂盒在信号增强方面通常不足,这是该方法的主要缺点。因此,具有高信号增强的灵敏、及时的现场 POCT 方法对于传染病的准确诊断至关重要。在这里,我们制备半胱胺金包覆的羧基铕螯合纳米粒子(Cys Au-EuNPs)介导的 POCT 用于检测 H5N1 禽流感病毒(AIV)。使用商业纳米粒子进行了比较。通过紫外可见分光光度计、荧光光谱仪、透射电子显微镜带有选区电子衍射(TEM-SAED)、傅里叶变换红外光谱(FTIR)和 zeta 电位分析证实了 Cys AuNPs、EuNPs 和 Cys Au-EuNPs 的光谱特征、表面形态、官能团、表面电荷和稳定性。粒径分布显示 Cys Au-EuNPs 的平均粒径约为 130 ± 0.66nm。Cys Au-EuNP 介导的 RDT(比色分析)和 FICT 试剂盒在不同滴度条件下对 H5N1 的检测限(LOD)分别为 10 HAU/mL 和 2.5 HAU/mL。所获得的 LOD 是商业纳米粒子缀合物的八倍。在紫外光照射下获得的 Cys Au-EuNPs 缀合物的光亮度(PL)稳定性约为 3%,与商业纳米粒子缀合物有很大差异。总体而言,开发的 Cys Au-EuNPs 介导的双模 POCT 试剂盒可作为开发传染病监测现场监测系统的有效纳米复合材料。