Generation of a recombinant antibody for sensitive detection of Pseudomonas aeruginosa
Gyu-Min Lim1, Joo-Kyung Kim1, Eun-Jung Kim2
1Interdisciplinary Program in Bioengineering, Seoul National University, Seoul, 08826, Republic of Korea.
Abstract:
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.
Insights
A new enzyme-linked immunosorbent assay (ELISA) detects Pseudomonas aeruginosa (P. aeruginosa) using a novel recombinant antibody against its V-antigen (PcrV). This method offers sensitive detection for controlling P. aeruginosa infections.
Area of Science:
- Microbiology
- Immunology
- Biotechnology
Background:
- Pseudomonas aeruginosa is a significant cause of hospital-acquired infections.
- Antibiotic resistance in P. aeruginosa complicates treatment and necessitates rapid detection methods.
- Controlling P. aeruginosa outbreaks requires accurate and sensitive diagnostic tools.
Purpose of the Study:
- To develop a sensitive enzyme-linked immunosorbent assay (ELISA) for detecting Pseudomonas aeruginosa.
- To produce and characterize a recombinant antibody (rAb) targeting the P. aeruginosa V-antigen (PcrV).
- To evaluate the efficacy of the rAb in a sandwich ELISA format for bacterial detection.
Main Methods:
- Production of a recombinant antibody (rAb) against P. aeruginosa PcrV in mammalian cells.
- Confirmation of rAb binding efficiency to P. aeruginosa.
- Development of a sandwich ELISA by pairing the rAb with a commercial anti-P. aeruginosa antibody.
- Determination of the limit of detection (LOD) for the developed ELISA.
Main Results:
- High-yield and high-purity production of the PcrV-specific rAb.
- Demonstrated binding efficiency of the rAb to P. aeruginosa.
- The sandwich ELISA showed an antigen-concentration-dependent response.
- Achieved a limit of detection of 230 CFU/mL for P. aeruginosa detection.
Conclusions:
- The developed rAb is effective for sensitive P. aeruginosa detection.
- The ELISA method shows promise for clinical diagnostics and point-of-care testing.
- This approach aids in controlling P. aeruginosa-related outbreaks through rapid identification.
More Related Videos
09:45Production of Monoclonal Antibodies Targeting Aminopeptidase N in the Porcine Intestinal Mucosal Epithelium
Published on: May 18, 2021
11:31Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa
Published on: June 30, 2016