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US 12,146,182 B2Granted
Methods and systems for the rapid detection of cronobacter using infectious agents
AssigneeLaboratory Corporation of America Holdings
InventorsMinh Mindy Bao Nguyen, Jose S. Gil, Stephen Erickson, Dwight Lyman Anderson, Jessica Stach
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Abstract
Disclosed herein are methods and systems for rapid detection of microorganisms such as Cronobacter 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 Cronobacter-specific bacteriophage, allows detection of a specific microorganism, such as Cronobacter spp. and an indicator signal may be amplified to optimize assay sensitivity.
Claims22
A method for detecting Cronobacter spp. in a sample comprising:
- incubating the sample with a cocktail composition comprising at least two different types of recombinant bacteriophages, wherein each of the at least two different types of the recombinant bacteriophages is a recombinant bacteriophage derived from a Cronobacter-specific bacteriophage comprising an indicator gene inserted into a late gene region of bacteriophage genome and, upstream of the indicator gene, a phage late gene promoter exogenous to the Cronobacter-specific bacteriophage, wherein the indicator gene is not contiguous with a gene encoding a structural bacteriophage protein and does not yield a fusion protein, and wherein expression of the indicator gene results in an indicator protein product;
- and,
- detecting the indicator protein product, the indicator protein product produced by the recombinant bacteriophage, wherein positive detection of the indicator protein product indicates that Cronobacter spp. is present in the sample,
- wherein the sample is a sample of infant formula, and
- wherein the at least two different types of the recombinant bacteriophages comprise at least one first recombinant bacteriophage derived from a podovirus Cronobacter-specific bacteriophage, and at least one second recombinant bacteriophage derived from a myovirus Cronobacter-specific bacteriophage.
2 more elements
Dependencies are inferred from the claim text.