Biological detection system and method

Inventors

Schofield, David Alexander

Assignees

Guild Associates Inc

Abstract

The present disclosure, according to some embodiments, relates to phage-based biological detection systems, compositions, and methods. In some embodiments, it relates to a detection system and method using phage binding and bacterial infection to detect the presence of a target molecule (e.g., a toxin). One detection system may include a genetically engineered phage that expresses a surface molecule able to bind a target molecule and/or target microorganism; a bacterium susceptible to infection by the phage; and a detection component able to determine whether the bacterium has been infected by the phage. Infection of a bacterium by a phage may be indicative of phage binding to the target molecule and/or target microorganism. One method may include placing a sample suspected of containing the target molecule and/or target microorganism with a binder; adding a genetically engineered phage having reporter genetic material and able to bind the target molecule and/or target microorganism; washing away unbound phage; releasing phage bound to the target molecule and/or target microorganism; infecting a bacterium with the released phage; and detecting the presence of any reporter genetic material in the bacterium. Reporter material in the bacterium may correlate with target molecule and/or target microorganism in the sample. In some embodiments, the disclosure relates to a detection system and method using phage comprising a reporter to infect a microorganism (e.g., Bacillus anthracis), wherein the reporter is selectively (e.g., only) detectable upon phage infection.

Core Innovation

The invention describes a phage-based detection system in which a genetically engineered phage captures a target molecule or target microorganism and then infects a susceptible bacterium so that a reporter becomes detectable only upon infection. The system includes genetically engineered phage displaying or expressing a binding molecule to enable binder-target binding, and the infected bacterium provides conditions for detection through reporter expression.

In the described examples, the system uses engineered M13 phage incorporating luxAB for E. coli detection, and engineered Wβ::luxAB phage for Bacillus detection. The detection behavior depends on spore germination, and the reporter signal timing occurs after infection-related progression.

The described engineered phage includes a reporter nucleic acid comprising at least one luxAB gene integrated into the lysogenic Wβ phage genome at a selected locus so that expression of the luxAB gene occurs in the infected Bacillus microorganism. A detector is provided to detect expression of the luxAB gene, with the disclosed workflow including binding/capture, washing to remove unbound phage, releasing bound phage, and infecting host bacteria prior to reporter detection.

Claims Coverage

The provided independent claim defines a kit with a specific engineered lysogenic Wβ phage carrying an integrated luxAB reporter nucleic acid and a detector configured to detect luxAB gene expression in an infected Bacillus microorganism. The inventive features are centered on three elements: lysogenic Wβ phage operable for Bacillus infection, reporter nucleic acid integration at specified wp loci, and detection of luxAB expression in infected Bacillus.

Overall, the claim coverage centers on using a genetically engineered lysogenic Wβ phage that infects Bacillus, carrying an integrated reporter nucleic acid with at least one luxAB gene inserted at wp39, wp40, or wp41 to replace a portion of the phage genome, together with a detector configured to detect luxAB expression after infection.

Stated Advantages

Documented Applications