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Last Updated: March 28, 2024

Claims for Patent: 5,238,813


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Summary for Patent: 5,238,813
Title: Diagnostic method for nephritis
Abstract:A diagnostic method is disclosed for determining the presence of renal disease or nephritis in association with systemic lupus erythematosus which comprises assaying for anti-glomerular binding reactivity in sera using a solid phase enzyme-linked immunoassay.
Inventor(s): Lefkowith; James B. (St. Louis, MO)
Assignee: Washington University (St. Louis, MO)
Application Number:07/896,851
Patent Claims:1. An in vitro diagnostic method for non-invasively determining anti-glomerular binding reactivity as distinguished from anti-DNA reactivity in the sera of a mouse or human host affected with SLE comprising:

a) digesting rat glomeruli with a mixture of collagenase and DNase type I enzymes,

b) binding said enzyme-digested glomeruli to a glass fiber carrier in a plurality of incubation wells,

c) incubating said enzyme-digested and carrier-bound glomeruli with a dilute buffered solution of from about 5% to 20% by weight blocking serum of a xenogeneic mammal in said wells,

d) reacting the thus-incubated glomeruli with serial dilutions of the sera of said mouse or human host for a predetermined period of time in said wells,

e) adding to said wells a detecting reagent of goat anti-mouse immunoglobulin or goat anti-human immunoglobulin linked to horseradish peroxidase,

f) allowing said detecting reagent to react for a predetermined period of time substantially similar to the time in step d,

g) adding to said wells a developing reagent of chloronaphthol,

h) observing the resulting color formation in said wells in which blue-black color formation indicates a positive diagnostic result of anti-glomular binding reactivity and yellow color formation indicates a negative diagnostic result, and in which said steps c through g are carried out in physiologically acceptable buffer at pH of about 7 to 8.

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