A.N. Bach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia.
Agricultural-Technological Institute, RUDN University, Miklukho-Maklaya Street 8/2, 117198 Moscow, Russia.
Sensors (Basel). 2019 Jan 4;19(1):153. doi: 10.3390/s19010153.
Lateral flow immunoassay (LFIA) is a convenient tool for rapid field-based control of various bacterial targets. However, for many applications, the detection limits obtained by LFIA are not sufficient. In this paper, we propose enlarging gold nanoparticles' (GNPs) size to develop a sensitive lateral flow immunoassay to detect . This bacterium is a quarantine organism that causes potato brown rot. We fabricated lateral flow test strips using gold nanoparticles (17.4 ± 1.0 nm) as a label and their conjugates with antibodies specific to . We proposed a signal enhancement in the test strips' test zone due to the tetrachloroauric (III) anion reduction on the GNP surface, and the increase in size of the gold nanoparticles on the test strips was approximately up to 100 nm, as confirmed by scanning electron microscopy. Overall, the gold enhancement approach decreased the detection limit of by 33 times, to as low as 3 × 10⁴ cells∙mL in the potato tuber extract. The achieved detection limit allows the diagnosis of latent infection in potato tubers. The developed approach based on gold enhancement does not complicate analyses and requires only 3 min. The developed assay together with the sample preparation and gold enlargement requires 15 min. Thus, the developed approach is promising for the development of lateral flow test strips and their subsequent introduction into diagnostic practice.
侧向流动免疫分析(LFIA)是一种用于快速现场控制各种细菌靶标的便捷工具。然而,对于许多应用,LFIA 获得的检测限还不够。在本文中,我们提出扩大金纳米颗粒(GNPs)的尺寸,开发一种用于检测的灵敏侧向流动免疫分析。这种细菌是一种检疫生物,会引起马铃薯褐腐病。我们使用金纳米颗粒(17.4 ± 1.0 nm)作为标记,并将其与针对的抗体偶联,制造了侧向流动测试条。我们提出了一种信号增强方法,即在金纳米颗粒表面还原四氯金(III)阴离子,从而增加测试条上金纳米颗粒的尺寸,大约增加了 100nm,这通过扫描电子显微镜得到了证实。总体而言,金增强方法将的检测限降低了 33 倍,在马铃薯块茎提取物中低至 3 × 10⁴个细胞·mL。实现的检测限允许对马铃薯块茎的潜伏感染进行诊断。基于金增强的开发方法不会使分析复杂化,只需要 3 分钟。开发的测定法与样品制备和金放大一起需要 15 分钟。因此,该方法有望用于开发侧向流动测试条,并随后将其引入诊断实践。