Method of detecting a pathogen using a virus
AssigneeHer Majesty the Queen in right of Canada, as represented by the Minister of National Defence of Her Majesty's Canadian Government
InventorsJohn W. Cherwonogrodzky, Kamil Lotfali
Life of this patent
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Abstract
A bacteriophage linked to an enzyme can replace an antibody in a system for detecting the presence of a bacteria in a sample. Specifically Brucella abortus (a pathogen which causes brucellosis in cattle) can be detected using Brucella bacteriophage for the virus, urease for the enzyme linked to the bacteriophage, m-maleimidobenzoyl-N-hydrosysuccimide ester as a coupling reagent, sera from mice immunized with Brucella bacteriophage for a detector antibody, urease conjugated to anti-mouse sheep antibody for an indicator, and urea with bromcresol purple as the substrate. The materials can be used in indirect (sandwich) or direct enzyme-linked viral assays (ELVirA).
Claims24
A method of detecting the presence of a bacteria in a sample comprising the steps of:
- (a) producing a conjugate comprising a wild type bacteriophage specific to said bacteria directly linked to an enzymatically active enzyme;
- (b) adding said conjugate to said sample to selectively bind said conjugate to said bacteria;
- and
- (c) detecting the presence of said bacteria by adding a substrate to said sample to generate a detectable change resulting from an enzymatic reaction between said enzyme and substrate.
A method of detecting the presence of a pathogenic bacteria in a sample comprising the steps of:
- (a) producing a conjugate comprising a bacteriophage specific to said pathogenic bacteria directly linked to an enzymatically active enzyme;
- (b) adding said conjugate to said sample to selectively bind said conjugate to said pathogenic bacteria;
- and
- (c) detecting the presence of said pathogenic bacteria by adding a substrate to said sample to generate a detectable change resulting from an enzymatic reaction between said enzyme and substrate.
A method of detecting a bacteria in a sample containing said bacteria and other microorganisms, said method comprising the steps of:
- (a) directly linking a bacteriophage specific to said bacteria to an enzymatically active enzyme to generate a bacteriophage-enzyme conjugate;
- (b) adding said bacteriophage-enzyme conjugate to said sample to selectively attach said bacteriophage-enzyme conjugate to said bacteria;
- (c) removing any unbound bacteriophage-enzyme conjugate and microorganism from said sample;
- and
- (d) adding a substrate to said sample, wherein said substrate is capable of promoting a detectable change with said enzyme to thereby enable detection of said bacteria attached to said bacteriophage-enzyme conjugate.
Dependencies are inferred from the claim text.