Interdisciplinary Program in Bioengineering, Seoul National University, Seoul, 08826, Republic of Korea.
BioMAX/N-Bio Institute, Institute of Bioengineering, Seoul National University, Seoul, 08826, Republic of Korea.
BMC Biotechnol. 2022 Aug 4;22(1):21. doi: 10.1186/s12896-022-00751-9.
Pseudomonas aeruginosa (P. aeruginosa) is a major pathogen that causes nosocomial infections and often exhibits antibiotic resistance. Therefore, the development of an accurate method for detecting P. aeruginosa is required to control P. aeruginosa-related outbreaks. In this study, we established an enzyme-linked immunosorbent assay method for the sensitive detection of three P. aeruginosa strains, UCBPP PA14, ATCC 27853, and multidrug-resistant ATCC BAA-2108. We produced a recombinant antibody (rAb) against P. aeruginosa V-antigen (PcrV), which is a needle tip protein of the type III secretion system of P. aeruginosa using mammalian cells with high yield and purity, and confirmed its P. aeruginosa binding efficiency. The rAb was paired with commercial anti-P. aeruginosa Ab for a sandwich ELISA, resulting in an antigen-concentration-dependent response with a limit of detection value of 230 CFU/mL. These results suggest that the rAb produced herein can be used for the sensitive detection of P. aeruginosa with a wide range of applications in clinical diagnosis and point-of-care testing.
铜绿假单胞菌（P. aeruginosa）是一种主要的病原体，可引起医院感染，并且经常表现出抗生素耐药性。因此，需要开发一种准确的方法来检测 P. aeruginosa，以控制与 P. aeruginosa 相关的爆发。在这项研究中，我们建立了一种酶联免疫吸附测定方法，用于灵敏检测三种铜绿假单胞菌菌株，UCBPP PA14、ATCC 27853 和多药耐药 ATCC BAA-2108。我们使用哺乳动物细胞高产率和高纯度地生产了针对铜绿假单胞菌 V 抗原（PcrV）的重组抗体（rAb），这是铜绿假单胞菌 III 型分泌系统的针状尖端蛋白，并证实了其与铜绿假单胞菌的结合效率。rAb 与商业抗铜绿假单胞菌 Ab 配对进行夹心 ELISA，导致抗原浓度依赖性反应，检测限为 230 CFU/mL。这些结果表明，本文中产生的 rAb 可用于灵敏检测铜绿假单胞菌，在临床诊断和即时检测中有广泛的应用。
