Diagnosis of pathogen infections through analysis of nitrite...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving oxidoreductase

Reexamination Certificate

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C435S029000, C435S383000

Reexamination Certificate

active

06673566

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The invention relates to a novel method for detecting tuberculosis and other infections in animals.
2. Description of the Prior Art
Cell-mediated immune responses are critical in the host defense against intracellular bacterial pathogens (Chan and Kaufmann, 1994, Immune mechanisms of protection, In Tuberculosis: Pathogenesis, protection, and control, B. R. Bloom, (ed.), American Society of Microbiology, Washington, D.C., pp. 389-415; Cheville et al., 1993, “Immune responses and protection against infection and abortion in cattle experimentally vaccinated with mutant strains of Brucella abortus,” American Journal of Veterinary Research 54:1,591-1,597; Chiodini, 1996. Immunology: Resistance to paratuberculosis, Veterinary Clinics of North America 12:313-342). A key component of this response is the clonal expansion of lymphocytes and the elaboration of cytokines that activate macrophages for the killing of bacteria located within the phagosomal compartment. Potent mediators of intra-phagosomal killing are reactive nitrogen intermediates (e.g., nitric oxide, NO) produced via the induction of inducible NO synthase (NOS), often as a sequalae to IFN-&ggr;, TNF-&agr;, or LPS stimulation (MacMicking et al., 1997, “Nitric oxide and macrophage function”, Annual Reviews of Immunology, 15:323-350; Kaufmann, 1999, “Cell-mediated immunity: Dealing a direct blow to pathogens”, Current Biology, 9:R97-99).
SUMMARY OF THE INVENTION
We have now discovered that peripheral blood mononuclear cells which have been isolated from an animal that is infected with a microbial pathogen produce nitric oxide in response to stimulation with antigens from that pathogen. Determination of nitric oxide production in cultures of peripheral blood mononuclear cells stimulated with a microbial pathogen's antigens may thus provide a specific indication of infection of the animal by that pathogen.
In accordance with this discovery, it is an object of this invention to provide an improved method for detecting an infection in an animal by a pathogenic microorganism.
Another object of this invention is to provide a method for detecting an infection in an animal which is specific for a particular microbial pathogen.
Yet another object of the invention is to provide a method for detecting Mycobacterium or Brucella infections in an animal.
Other objects and advantages of this invention will become readily apparent from the ensuing description.


REFERENCES:
patent: 3634198 (1972-01-01), Truhan
patent: 4631255 (1986-12-01), Takino et al.
patent: 5910421 (1999-06-01), Small, Jr. et al.
patent: 6326357 (2001-12-01), Phillips et al.

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