Bioluminescence-based Reporter Phage Detection Assay

As a world-leading phage detection service provider, Creative Biolabs focuses on providing customized phage detection services to customers around the world. Relying on our state-of-the-art manufacturing facilities and decades of experience, we have the confidence to customize high-quality bioluminescence-based pathogen detection services to meet your unique needs.

Overview of Bioluminescence-based Reporter Phage

As we all know, Phages are a class of viruses that specialize in infecting bacteria. They are ideal for developing highly specific diagnostic analysis systems. The construction of bioluminescent reporter phages by integrating luciferase genes into viral genomes is emerging as an attractive strategy for pathogen detection. These phages specifically infect host cells and transduce a heterologous luciferase gene, the activity of which can be detected to indicate the presence of target cells. This strategy is superior to any other molecular detection method because gene expression and light emission depend on the active metabolism of bacterial cells, and only living cells generate signals.

Principle and Workflow

The construction of bioluminescent reporter phages follows a crucial principle: the reporter gene is integrated into the phage genome. A key step in bioluminescence-based reporter phage engineering is to identify a highly expressed phage protein so that its promoter can be used to drive the expression of the introduced gene. The standard approach is to add reporter bricks after the major capsid protein open reading frame (ORF) to obtain a highly sensitive luminescent phage. Alternative strategies are needed when standard methods do not yield target luminescent phages. In this strategy, a reporter operon consisting of a promoter, reporter gene, and terminator is inserted into several different phage genome locations. Recombinant phage carrying a luminescent reporter operon emits a detectable signal when the phage interacts with target bacteria.

Fig.1 Strategy of Reporter Operon cloning leading to the isolation of luminescent reporter phages. (Zelcbuch, 2021)

Advantages

• Highly specific - phages are naturally bacterial-specific.
• Highly sensitive - most samples are unlikely to have background activity that masks or produces false positive signals in bioluminescence measurements.
• Rapid - this approach has enabled exciting detection times that are comparable to molecule-based approaches.
• Simplicity - the procedure is simple and easy to perform and requires a minimum of equipment and training to use.
• Cost-effective - phage reagents are inexpensive compared to molecular diagnostics.

Services at Creative Biolabs

At Creative Biolabs, our expert scientists combine efficient enrichment and immobilization technology with bioluminescent phage detection, which provides a powerful tool for the specific detection of bacterial pathogens. Pre-enrichment of target bacteria with magnetic separation followed by detection with bioluminescent reporter phage provides more reliable results. We are confident to provide comprehensive bioluminescence-based reporter phage detection services, from experimental protocol design to final experimental results analysis. If you have any difficulties with your phage-based pathogen detection project, please feel free to contact us for professional assistance.

Reference:

1. Zelcbuch, L.; et al. Luminescent Phage-Based Detection of Klebsiella pneumoniae: From Engineering to Diagnostics. Pharmaceuticals. 2021, 14(4): 347.