Bacteriophage technology

Bacteriophage can be described as bacteria-specific viruses. Unlike viruses, bacteriophages are safe to humans, animals, and anything that is not bacteria. Through evolution, bacteriophages are capable of efficiently and effectively replicating in the presence of specific bacteria.

Phage amplification
MicroPhage's technology represents a well-defined and novel approach to rapidly identifying pathogens. Specific bacteriophage are introduced to the processed sample, where they find, infect, and amplify their target pathogen(s) providing a very specific, highly amplified surrogate marker for rapid detection.

Bacteriophage selectively infect targeted bacteria and rapidly multiply. The system can generate as much as a 5-log amplification of signal. This allows for reduced incubation times, resulting in complete assay times of one to four hours compared with traditional culture assays of 24 to 48 hours.

Compared to the replication time of bacteria, bacteriophage multiply very rapidly within their target bacteria. In a short time, tens to thousands of progeny phages are produced. Pressure increases within the cell or singular cellular burst events cause the release of the progeny phages. This increase in bacteriophage concentration indicates the presence of the target bacteria.

In antibiotic tests, the bacteria are exposed to the target antibiotic and the bacteriophage. Since bacteriophage amplification requires a viable, replicating bacterial host, susceptible organisms cannot support phage amplification and can be differentiated from resistant bacteria that do support bacteriophage amplification.

The phage life cycle
MicroPhage uses the well-defined life cycle of lytic bacteriophage to detect target bacteria.

Adsorption

Bacteriophage quickly find specific receptors on the surface of the target bacteria and attach in a two-step process. First, the phage tail fibers create a low-affinity bind to the outer membrane of the bacterium. Second, small tail fibers extend from the baseplate of the bacteriophage to form a high-affinity, irreversible bond to the bacterium. Enzymes located on this baseplate allow the softening of the bacterium's membrane.

Injection

Once the phage creates an irreversible bond to the bacterium and the enzyme has sufficiently compromised the membrane, the phage injects its nucleic acid into the bacterium.

Replication

Following nucleic acid injection, the phage is replicated into hundreds to thousands of progeny bacteriophage. The parent bacteriophage's nucleic acid inserts itself into the DNA replication process of the bacterium host, halting further bacteria growth and forcing it to produce proteins that will assemble and become bacteriophage.

Lysis

Depending on the bacteriophage, an additional enzyme is formed following, or in concert with, bacteriophage replication. This step produces holing or lysine, which thins the bacterial membrane and allows the progeny phage to break into the open environment.

This step occurs for each successfully infected bacterium. With high concentrations of bacteria, lysis will occur almost in unison for all phage-infected bacteria.