Colorimetry-based Reporter Phage Detection Assay

Creative Biolabs is a global leading innovative biotechnology company focused on phage-based bacterial pathogen detection research. After decades of accumulation, we have accumulated rich experience in the phage detection field. Our vision is to help customers around the world achieve breakthrough progress in phage-related projects.

Background of Colorimetric Detection

Bacterial infectious diseases are a major cause of death worldwide. Accurate and early detection of bacterial pathogens facilitates effective measures to reduce outbreaks of pathogens. However, many methods for identifying pathogenic bacteria take more than ten hours from sampling to results. Therefore, developing fast and reliable methods to detect bacteria in environmental, food and water matrices is an urgent task. In recent years, colorimetry-based reporter phage detection has gradually emerged as an attractive method for the rapid detection of bacterial pathogens.

Basic Principle and Workflow

Phages can specifically infect target host cells and rapidly propagate in the host before lysing the cells. The reporter gene, when expressed, provides an efficient method for detecting bacteria using standard colorimetric, fluorescent, or luminescent signals. Because phages are special viruses, their genes are not expressed until the host is infected, thus confirming the existence of viable host bacteria. A canonical example is the design of phage T7 to overexpress a carbohydrate-binding module (CBM)-tagged alkaline phosphatase reporter gene upon infection of E. coli in a model system. Following phage infection of the host bacterium, the CBM-containing reporter enzyme is expressed. CBM fusion tags enable the immobilization of reporter genes on magnetic cellulose particles. Particles with immobilized enzymes are isolated and concentrated. After the addition of substrate, the resulting color was assessed by a spectrophotometer or visual inspection.

Fig. 1 Phage-based pathogen detection.¹

Advantages

Colorimetry-based reporter phage detection is a colorimetric bacterial detection method. The method is fast and cost-effective. Furthermore, colorimetric assays that do not rely on advanced instruments and skilled operators offer promising tools for the detection of bacterial pathogens. Typically, this assay does not require the isolation of bacteria from food matrices, thus reducing the complexity of the assay and can be used to detect bacteria in complex samples. Most importantly, the assay can be completed in less than an hour and is stable in different media such as seawater and human serum. Therefore, colorimetry-based detection has received increasing attention in recent years.

At Creative Biolabs, our customer-oriented team of seasoned experts has earned a reputation for high-quality service. As an expert in the field of phage detection, our experienced technicians will work with you to customize a reasonable solution to facilitate the progress of your project. Please feel free to contact us for more information and detailed quotation.

Reference:

1. Meile, Susanne, et al. "Reporter phage-based detection of bacterial pathogens: design guidelines and recent developments." Viruses 12.9 (2020): 944. Under Open Access license CC BY 4.0, without modification.