Bacteriophages Isolation and Efficacy Testing

Abstract

Bacteriophages (phages) are viruses that infect bacteria and are the most abundant biological entity on the planet. Phages have gained popularity as an alternative to antibiotics due to their specificity and ability to efficiently lyse antimicrobial resistant bacterial pathogens. Before using phages, they must be isolated from the environment and tested to ensure purity and lytic ability against various hosts. This protocol walks through the entire multi-day procedure of enriching and processing raw environmental samples (seawater, primary sludge, and soil), testing for lytic activity, selecting and picking potential phage plaques, verifying phage purity, and finally, propagation (liquid and solid) of phages to obtain high-titer crude phage lysates.

Keywords: Bacteriophage; Double-layer agar methods; Isolation; Lytic phage; Lytic plaques; Propagation; Purification.