Composition, formulae, devices and methods for control of specificity and inclusivity of microorganisms containing closely related antigen epitopes

a technology of antigen epitopes and microorganisms, which is applied in the field of compositions, formulae, devices, and methods for detecting and identifying microorganisms, can solve the problems of death complications, severe abdominal cramps, renal failure,

Inactive Publication Date: 2002-09-12
BIOCONTROL SYST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

0026] These and other aspects of the present invention will become evident upon reference to the following detailed description and examples. In addition, the various references set forth be

Problems solved by technology

EHEC is particularly virulent and can trigger deadly complications, including severe abdominal cramps and acute renal failure in children as well as cardiovascular and central nervous system problems.
One particularly troublesome aspect of Listeria control in foods is that Listeria can grow at temperatures as low as -0.4.degree. C. and as high as 44.degree. C.
While taking much less time than the cultural methods, these rapid tests still require skilled technical training, a functional laboratory, and specialized equipment.
These tests generally take a total of two or more days, including considerable hands-on time.
Campylobacter detection methodology to date is technically intensive requiring fastidious media and environmental conditions, in addition to well-trained analysts.
These challenges are the need to detect all of the strains of a pathogenic microorganism in the presence of numerous antigenically related

Method used

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Examples

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example 1

[0051] Nine strains of Salmonella were identified which were non-reactive in a visual immunoprecipitate assay. The strains were reported to produce excessive levels of surface antigen. It was hypothesized that growth of the strains in the inventive media would eliminate or significantly reduce the expression of surface O group antigen epitopes, thereby allowing detection by highly specific monoclonal antibodies directed against the core region of lypopolysaccharide contained in the visual immunoprecipitate assay device.

[0052] The organisms were grown in the inventive medium, followed by extraction with 0.1% SDS at 100.degree. C. for ten minutes. Strong reactivity was demonstrated. The inventive media formulation was a commercially available formulation of tryptic soy broth containing the following ingredients supplemented with 0.5 mM (0.01%) 2,4-dinitrophenol.

1 Trypticase Soy Broth Pancreatic Digest of Casein 17.0 g Papaic Digest of Soybean Meal 3.0 g Sodium Chloride 5.0 g Dipotassi...

example 2

[0053] Two strains of Salmonella, serogroups A and B, were determined to be weakly reactive in a monoclonal antibody based visual immunoprecipitate assay and an enzyme linked immunoassay in the presence of related competitive microorganisms. These strains were incubated in the inventive medium in a 1000 fold excess of competitive bacteria, and found to be highly reactive. The inventive media formulation was a commercially available formulation of buffered peptone water supplemented with 0.5 mM (0.01%) 2,4-dinitrophenol.

2 Buffered Peptone Pancreatic Digest of Gelatin 10.0 g Sodium Chloride 5.0 g Disodium Phosphate 3.5 g Monopotassium Phosphate 1.5 g Distilled Water 1000 ml

example 3

[0054] A strain of E. Coli 0157:H7 was found to be weakly reactive in a polyclonal antibody based assay. The strain was grown in the inventive medium and the sensitivity was improved by 100 fold. The media was a modified tryptic soy broth with 20 mg / ml novobiocin supplemented with 0.5 mM (0.01%) 2,4-dinitrophenol.

3 Modified Tryptic Soy Broth Bacto Tryptone 17.0 g Bacto Soytone 3.0 g Sodium Chloride 5.0 g Dipotassium Phosphate 4.0 g Bile Salts No. 3 1.5 g Bacto Dextrose 2.5 g

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Abstract

Compositions, formulae, devices and methods for the detection of target microorganisms, such as by visual immunoprecipitate assay, enzyme linked immunoassay, chemiluminescence, immunoblotting, or similar detection technology, wherein detection requires the discrimination among closely related genera, species and strains of antigenically related microorganisms based on immunological reactivity of a highly conserved antigen epitopes with a reagent system comprised of an antibody linked to a detecting reagent. The invention permits a detectable event to occur by exposing inaccessible but highly conserved and specific antigen epitopes to the detecting reagent. Exposure of such antigen epitopes without inactivating microbial metabolism allows for specific detection.

Description

CROSS-REFERENCE TO RELATED APPLICATION[0001] This application is a continuation of U.S. patent application Ser. No. 09 / 374,560, filed Aug. 13, 1999, now allowed; which application claims priority from U.S. Provisional Application No. 60,096,566, filed Aug. 14, 1998; which applications are incorporated herein by reference in their entirety.TECHNICAL FIELD[0002] The present invention relates generally to compositions, formulae, devices, and methods for detecting and identifying microorganisms and, more particularly, to compositions, formulae, devices, and methods for detecting microorganisms by exposing highly conserved antigenic sites of target microorganisms without inactivating the microorganisms' ability to grow to sufficient levels to be detected by an antibody linked detection system.BACKGROUND OF THE INVENTION[0003] Microbial diseases have long been a major health concern worldwide. Significant increase in the frequency and severity of outbreaks have occurred throughout the wor...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12N1/20C12Q1/04C12R1/01G01N33/569
CPCC12Q1/04C12Q1/045G01N33/56916G01N33/56922G01N2333/205G01N2333/245G01N2333/255
Inventor FELDSINE, PHILIP T.KERR, DAVID E.ZHU, PINGMUI, LINDA
Owner BIOCONTROL SYST
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