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

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...

AI Technical Summary

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 organisms, with a low tolerance for false positive results and a very low, preferably zero, tolerance for false negatives.

The second challenge is the physical size and heterogeneity of the microorganism itself.

The ability to isolate and bind to antibodies reacting with these highly conserved antigen epitopes is difficult, however, because they can be sterically hindered by O-antigen polysaccharide chains.

They are generally inaccessible because of a phenomenon known as steric interference.

Although, aggressive treatments are available which will expose interior antigen epitopes, these treatments destroy cell viability and in many cases disrupt cellular integrity completely.

The significant shortcoming of these extractions is that they result in death of the microorganism.

Therefore, if the cell population had not reached a sufficiently detectable level prior to inactivation, a negative determination will result.