Template reporter bacteriophage platform and multiple bacterial detection assays based thereon
AssigneeUniversity of Wyoming
InventorLawrence Goodridge
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Abstract
The invention is a method for the development of assays for the simultaneous detection of multiple bacteria. A bacteria of interest is selected. A host bacteria containing plasmid DNA from a T even bacteriophage that infects the bacteria of interest is infected with T4 reporter bacteriophage. After infection, the progeny bacteriophage are plating onto the bacteria of interest. The invention also includes single-tube, fast and sensitive assays which utilize the novel method.
Claims14
A method for the simultaneous development of multiple bacterial detection assays, comprising the steps of:
- (a) selecting a bacteria of interest;
- (b) selecting a T even bacteriophage that infects said bacteria of interest;
- (c) providing a T4 reporter bacteriophage;
- (d) providing a bacterial host that contains host range cassette plasmid DNA cloned from the selected T even bacteriophage;
- (e) infecting said bacterial host with said T4 reporter bacteriophage;
- and
- (f) plating progeny bacteriophage on said bacterial host of interest.
A method as defined in claim 2, wherein the bacteria of interest is selected from the group consisting of Escherichia coli, Salmonella spp., Shigella spp., Yersinia spp., Aeromonas spp., Burkholderia spp., Pseudomonas spp., Acinebacter spp., Vibrio spp., Klebsiella, spp., Citrobacter spp., Proteus spp., and Serratia spp.
An assay for detecting the presence of viable target bacteria in a sample, comprising:
- (a) a container for the sample;
- (b) media in the container which supports growth of the target bacteria;
- (c) specific separation beads in the container to concentrate the target bacteria;
- (d) a sampling device within the container to rapidly obtain the sample to be tested;
- (e) a reporter phage in the container capable of infecting a viable target bacterium in the sample which upon incubation produces the reporter;
- and
- (f) a detector substrate released into the media which produces a signal in the presence of the reporter.
Dependencies are inferred from the claim text.