Phage Isolation and Identification Services
BackgroundService ProcedureOur AdvantagesCase StudyFAQs
Background
Bacteriophages (phages) are viruses that specifically infect and lyse bacterial cells. With the increasing prevalence of antibiotic-resistant bacterial strains, phages have re-emerged as a promising alternative or complementary strategy in both therapeutic and industrial microbiology. As a result, phage isolation and identification services have become critical components in the development of phage-based applications.
Phage isolation involves the targeted extraction of bacteriophages from environmental samples such as soil, water, wastewater, or host-associated microbiota. These environments are typically rich in microbial diversity and serve as natural reservoirs for bacteriophages. Isolation is performed using specific bacterial strains as hosts, allowing selective enrichment of phages with lytic activity against target bacteria.
After successful isolation, phage identification is performed to characterize the biological and genetic properties of the phages. This process includes assessment of host range, plaque morphology, genome sequencing and phylogenetic analysis. In certain cases, transmission electron microscopy is used to determine phage morphology and taxonomic classification. This comprehensive characterization ensures that the selected phages are both effective against the intended bacterial targets and meet safety and regulatory standards, particularly in clinical or agricultural contexts.
Figure 1. Schematic illustration of the phage isolation, purification, and preservation. (Adapted from Jaglan et al., 2022)
Creative BioMart Microbe offers a comprehensive suite of phage services, including phage isolation and identification, supporting a wide range of applications, including the development of personalized phage therapies, targeted biocontrol in food and agriculture, and microbiome modulation strategies. Contact us for more information!
Service Procedure
Service Details
Natural Phage Discovery and Isolation Services
We begin by obtaining environmental samples-such as wastewater, soil, or effluent-to identify bacteriophages capable of infecting your target bacterial strains. Through careful enrichment and plaque assays, we isolate and amplify phages with the desired host specificity.
Phage Identification Services
Comprehensive characterization is critical to understanding the therapeutic potential of isolated phages. We perform genomic sequencing, host range determination, lytic activity assays, and stability studies to provide a detailed profile of each phage.
Service Offering
Delivering effective phage-based solutions begins with precision isolation and identification. At Creative BioMart Microbe, our phage isolation and identification services provide a robust, end-to-end workflow designed to recover, purify, and genetically characterize high-quality bacteriophages. Whether the goal is therapeutic development, microbiome modulation, or biocontrol applications, each stage of the process must be scientifically validated, reproducible, and tailored to the microbial target. Explore the details of our services to isolate and identify phages with confidence and regulatory farsightedness.
Sample Collection
Collect environmental or clinical samples rich in phage biodiversity—such as wastewater, soil, or infected tissue—using sterile, quality-controlled protocols to maximize viable phage recovery.
Enrichment Culture
Co-incubate target bacteria with collected samples under optimized conditions to amplify lytic phages and enhance detection sensitivity.
Filtration & Clarification
Filter samples through 0.22 µm membranes to remove bacterial cells and debris, isolating bacteriophages for downstream analysis.
Plaque Assay (Phage Hunting)
Perform double-layer agar plaque assays to detect phage-induced bacterial lysis, visually confirming the presence of active bacteriophages.
Phage Purification
Isolate individual plaques and perform serial plating to ensure clonal purity and prepare individual phages for characterization.
Phage Titration
Quantify phage concentrations by calculating plaque-forming units per milliliter (PFU/mL), establishing standardized potency metrics.
Phage Characterization
Analyze phage morphology and host range by electron microscopy and infectivity profiling to determine application suitability.
Genomic DNA Extraction
Extract high-quality, contaminant-free phage DNA using proprietary methods optimized for downstream sequencing and diagnostics.
Whole Genome Sequencing
Sequencing of complete phage genomes by next-generation sequencing (NGS) to assess safety, therapeutic potential, and genetic characteristics.
Bioinformatic Analysis & Library Matching
Compare phage genomes against curated databases to classify phages, predict functionalities, and ensure compliance with regulatory standards.
Our Advantages
- Expertise: Leverages specialized experience in phage biology, therapeutic development, and microbiology to deliver accurate, reliable, and innovative research services.
- Customization: Designs and implements customized phage isolation and identification strategies, ensuring that solutions are perfectly aligned with each client's unique scientific goals and challenges.
- State-of-the-Art Facilities: Operates world-class laboratories equipped with advanced phage research technologies, high-throughput screening systems, and robust bioinformatics platforms for maximum innovation.
- Regulatory Compliance: Maintains strict adherence to international quality standards, including GMP, GLP, and ISO certifications, to ensure safety, efficacy, and ethical compliance.
Case Study
Case Study 1: Isolation of phages for phage therapy: comparing spot tests and EOP for host range and efficacy.
Future phage therapy will not be "on demand" due to the high cost and regulatory requirements of clinical trials. Although numerous phages can be isolated for each bacterial strain, only highly virulent phages with broad host ranges and large burst sizes are economically viable for development. Efficiency of plating (EOP) is a more reliable selection method than spot testing, which often misrepresents virulence. In this study, phages isolated from the ECOR (E. coli reference) collection showed efficacy against several ESBL (extended-spectrum beta-lactamases) E. coli strains, demonstrating the potential of non-pathogenic screening strains. Broader screening and EOP analysis of more strains could identify even more effective phages. In addition, understanding bacterial resistance mechanisms—particularly those encoded by prophages such as E. coli phage P2, which can inhibit virulent phage infections—is essential. In conclusion, EOP assays are more reliable for building therapeutic phage libraries.
Figure 2. Transmission electron micrographs of the six phages in this study. (Khan Mirzaei and Nilsson, 2015)
Case Study 2: Characterization of crAss-like phage isolates highlights Crassvirales genetic heterogeneity and worldwide distribution.
CrAssvirales (crAss-like phages), abundant in the human gut and proposed as indicators of fecal contamination, are poorly understood due to the limited number of cultured isolates. This study reports the isolation and genetic characterization of 25 new crAss-like phages (crAssBcn) infecting Bacteroides intestinalis. These phages, all podovirus-like, fall into six species and show genomic diversity distinct from previously known strains such as ΦCrAss001. Using qPCR assays and enrichment cultures from wastewater samples, the researchers identified B. intestinalis as the optimal host for propagation, enabling successful isolation. The discovery significantly expands the known diversity of crAss-like phage and raises the question of which variant is best suited as a fecal marker.
Figure 3. Comparison of the six representative crAssBcn phages of each species and ΦCrAss001. Electron micrographs and CDS alignment of the genomes of six crAssBcn phages representative of the six species (groups I-VI). (Ramos-Barbero et al., 2023)
FAQs
Q: What types of samples do you accept for phage isolation?
A: We accept a variety of environmental samples, including sewage, soil, wastewater, and clinical isolates, depending on the project requirements.
Q: How do you determine the host range of isolated phages?
A: We use spot testing, plaque assays, and Efficiency of Plating (EOP) analysis against a panel of bacterial strains to define the host range and specificity. This ensures accurate assessment of therapeutic or biocontrol potential.
Q: Are your services available for clinical applications?
A: Currently, our services are designed for research and development purposes. Clinical applications are subject to regulatory approvals and are not part of our standard offerings.
Q: How long does the entire phage development process take?
A: A: The timeline varies depending on the complexity of the project, but typically ranges from 8 to 12 weeks from consultation to final report.
Q: What makes your phage isolation services stand out?
A: Our services are fully integrated—from sample processing to host range profiling and genetic analysis. We combine high-throughput screening with regulatory foresight to deliver research- and application-ready phages.
References:
- Gill JJ, Hyman P. Phage choice, isolation, and preparation for phage therapy. Current Pharmaceutical Biotechnology. 11(1):2-14. doi:10.2174/138920110790725311
- Hyman P. Phages for phage therapy: isolation, characterization, and host range breadth. Pharmaceuticals. 2019;12(1):35. doi:10.3390/ph12010035
- Jaglan AB, Anand T, Verma R, et al. Tracking the phage trends: A comprehensive review of applications in therapy and food production. Front Microbiol. 2022;13:993990. doi:10.3389/fmicb.2022.993990
- Khan Mirzaei M, Nilsson AS. Isolation of phages for phage therapy: a comparison of spot tests and efficiency of plating analyses for determination of host range and efficacy. Schuch R, ed. PLoS ONE. 2015;10(3):e0118557. doi:10.1371/journal.pone.0118557
- Ramos-Barbero MD, Gómez-Gómez C, Sala-Comorera L, et al. Characterization of crAss-like phage isolates highlights Crassvirales genetic heterogeneity and worldwide distribution. Nat Commun. 2023;14(1):4295. doi:10.1038/s41467-023-40098-z
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