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Lateral flow assay-based bacterial detection using engineered cell wall binding domains of a phage endolysin
Minsuk Kong1, Joong Ho Shin2, Sunggi Heu3
1Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Center for Food and Bioconvergence, Research Institute of Agriculture and Life Sciences, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 08826, Republic of Korea.
A new lateral flow assay uses engineered phage cell wall binding domains (CBDs) for rapid, cost-effective bacterial detection. This biosensor offers a promising alternative to traditional antibody-based methods for diagnosing Bacillus cereus (B. cereus).
Area of Science:
- Biotechnology
- Biosensor Development
- Bacteriology
Background:
- Cost-effective and efficient bacterial detection assays are crucial for diagnostics, especially in resource-limited areas.
- Traditional antibody production for bacterial capture is expensive and time-consuming, hindering widespread application.
Purpose of the Study:
- To develop a novel, cost-effective lateral flow assay for detecting the pathogenic bacterium Bacillus cereus (B. cereus).
- To utilize engineered phage cell wall binding domains (CBDs) as a recognition element for improved bacterial detection.
Main Methods:
- A nitrocellulose-based lateral flow assay was designed using phage-derived cell wall binding domains (CBDs).
- Engineered CBDs, tagged with cysteine-glutathione-S-transferase or maltose-binding protein, were produced in Escherichia coli (E. coli).
- Colloidal gold nanoparticles were employed as a colorimetric signal for B. cereus detection.
Main Results:
- The developed assay achieved a sensitivity of 1x10^4 CFU/mL for B. cereus detection.
- The entire assay procedure was completed within 20 minutes.
- The CBD-based assay demonstrated superior performance compared to antibody-based methods, with no significant cross-reactivity observed.
Conclusions:
- Engineered phage CBDs show significant promise as recognition elements in lateral flow assays for whole bacterial cell detection.
- This proof-of-concept study highlights the potential of these assays as simple, rapid, and cost-effective biosensors.
- The developed method offers a viable alternative for bacterial diagnostics in various settings, including resource-poor environments.
