Methods and Systems for the Rapid Detection of Microorganisms Using Recombinant Infectious Agents to Express an Indicator Subunit
AssigneeLaboratory Corporation of America Holdings
InventorsStephen E. Erickson, Jose S. Gil, Wendy Hahn, Dwight L. Anderson
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Abstract
Disclosed herein are methods and systems for rapid detection of microorganisms such as bacteria in a sample. A genetically modified bacteriophage is also disclosed which comprises an indicator gene encoding one subunit of an indicator protein. The specificity of the bacteriophage allows detection of a particular bacteria of interest and an indicator signal may be amplified to optimize assay sensitivity.
Claims15
A recombinant indicator bacteriophage comprising an indicator gene inserted into the bacteriophage genome, wherein the indicator gene encodes a peptide or polypeptide subunit of an indicator protein.
A method of preparing a recombinant indicator bacteriophage comprising:
- selecting a wild-type bacteriophage that specifically infects a target pathogenic bacterium;
- preparing a homologous recombination plasmid/vector comprising an indicator gene;
- transforming the homologous recombination plasmid/vector into target pathogenic bacteria;
- infecting the transformed target pathogenic bacteria with the selected wild-type bacteriophage, thereby allowing homologous recombination to occur between the plasmid/vector and the bacteriophage genome;
- and,
- isolating a particular clone of recombinant bacteriophage.
A kit for detecting a particular bacteria of interest comprising a recombinant indicator bacteriophage, comprising an indicator gene encoding a first peptide or polypeptide subunit of an indicator protein.
A system for detecting a particular bacteria of interest comprising a recombinant indicator bacteriophage, comprising an indicator gene encoding a peptide or polypeptide subunit of an indicator protein.
Dependencies are inferred from the claim text.