Methods and systems for the rapid detection of Listeria using infectious agents
AssigneeLaboratory Corporation of America Holdings
InventorsStephen Erickson, Jose S. Gil, Matthew J. Brown, Minh Mindy Bao Nguyen, Wendy Hahn
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Abstract
Disclosed herein are methods and systems for rapid detection of microorganisms such as Listeria spp. in a sample. A genetically modified bacteriophage is also disclosed which comprises an indicator gene in the late gene region. The specificity of the bacteriophage, such as Listeria-specific bacteriophage, allows detection of a specific microorganism, such as Listeria spp. and an indicator signal may be amplified to optimize assay sensitivity.
Claims25
A recombinant bacteriophage (i) comprising a bacteriophage LPJP1 having a phage genome of at least 200 kbp and (ii) comprising a genetic construct comprising an indicator gene, wherein the recombinant bacteriophage specifically infects Listeria grayi.
A method of preparing a recombinant indicator bacteriophage comprising:
- selecting a wild-type bacteriophage LPJP1 having a phage genome of at least 200 kbp that specifically infects Listeria grayi;
- 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 wild-type bacteriophage LPJP1, thereby allowing homologous recombination to occur between the plasmid/vector and the bacteriophage genome;
- and
- isolating a particular clone of recombinant bacteriophage comprising the indicator gene.
Dependencies are inferred from the claim text.