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Method and apparatus for distinguishing Crohn's disease from ulcerative colitis and other gastrointestinal diseases by detecting the presence of fecal antibodies to Saccharomyces cerevisiae

a technology of saccharomyces cerevisiae and fecal antibodies, which is applied in the field of method and apparatus for the differentiation of crohn's disease from other gastrointestinal illnesses, can solve the problems of not being able to achieve rapid differential diagnosis and serum antibodies may not be accurate indicators of proper diagnosis

Inactive Publication Date: 2005-06-23
TECH LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The patent describes a method for testing fecal samples to diagnose Crohn's disease. The method involves measuring the amount of anti-Saccharomyces cerevisiae antibodies present in the sample. This test can help to differentiate Crohn's disease from other gastrointestinal illnesses. An assay is provided to determine the concentration of these antibodies, and a diagnostic assay is also included to compare the level of antibodies in the sample to a reference value for healthy control subjects. The invention provides a simple and effective tool for diagnosing Crohn's disease."

Problems solved by technology

These methods, however, do not permit quick differential diagnosis as each may require years for a diagnosis to be confirmed.
Further, each method utilizes only serum antibodies that are not required to cross the intestinal wall and the serum antibodies may not be accurate indicator for the proper diagnosis.

Method used

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  • Method and apparatus for distinguishing Crohn's disease from ulcerative colitis and other gastrointestinal diseases by detecting the presence of fecal antibodies to Saccharomyces cerevisiae
  • Method and apparatus for distinguishing Crohn's disease from ulcerative colitis and other gastrointestinal diseases by detecting the presence of fecal antibodies to Saccharomyces cerevisiae

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0020] In this example using an ELISA method, a fecal sample was obtained and serially diluted 20 fold. 100 μl of the diluted sample was added to a test well of a microassay plate coated with extract of Saccharomyces cerevisiae and purified mannan. The sample then was incubated at 37° C. to allow antibodies to Saccharomyces cerevisiae to bind to the extract of Saccharomyces cerevisiae. Following incubation, anti-human Ig polyclonal antibodies coupled to horseradish peroxidase enzyme (conjugate) were added to the test well and allowed to bind to captured ASCA. Unbound conjugate then was washed from the well and one component substrate (tetra-methyl-benzidene and hydrogen peroxide) was added for color development. Following the substrate incubation, 0.1M sulfuric acid was added to quench the reaction and the optical density (OD) was obtained spectrophotometrically at 450 nm using a single wavelength spectrophotometer.

[0021] The method described above was used in a clinical study to t...

example 2

[0030] In this example, the sensitivity of the fecal ASCA test was determined using serial two fold dilutions of highly purified ASCA antibodies. For the analysis, standard curves were generated using the kit dilutent. The test was consistently positive at a concentration of 0.62 μg / mL as determined by a cutoff absorbency value of ≧0.200. Individual results are shown below in Table 10. The standard curves are shown in FIG. 1.

TABLE 10Standard curves generated using purified ASCA antibodiesPurifiedASCAAntibodies(μg / mL)Test 1Test 2MeanStd Dev5.001.7021.8561.7790.1082.501.1171.0991.1080.0121.250.6340.6240.6290.0070.620.3030.3290.3160.0180.310.1910.1640.1770.0190.160.1150.1130.1140.0010.080.0900.0770.0830.0090.040.0630.0650.0640.001

example 3

[0031] In this example, tests were conducted to determine what type of immunoglobulins (antibodies) were present in a fecal sample and in serum. The immunglobulin typing was done for human IgA, human IgAsec, human IgD, human IgM, and human IgG. The immunoglobulin typing was done on a fecal sample from 6 Crohn's disease patients and 2 ulcerative colitis and on a serum control sample using pre-absorbed Ig-type specific conjugates. The serum control sample was obtained from a patient with a confirmed allergy to Saccharomyces cerevisiae.

[0032] Of the Crohn's disease patients, 5 patients exhibited a response to IgA and IgAsec, 4 patients exhibited a response to IgM and a single patient exhibited a response to IgG. Of the 2 ulcerative colitis patients, a single patient reacted with the Ig conjugate. The serum control only exhibited a response to individual immunoglobulins IgM and IgG. A response to IgA and IgAsec occurred the fecal samples but not in the control serum sample. A summary o...

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Abstract

A method and apparatus for the differentiation of Crohn's disease from other gastrointestinal illnesses, such as ulcerative colitis and irritable bowel syndrome, using the presence of fecal anti-Saccharomyces cerevisiae antibodies (ASCA) as a marker for Crohn's disease are provided. The apparatus includes an enzyme-linked immunoassay or other immunoassay that utilizes antibodies specific to human immunoglobins for the measurement of total endogenous ASCA in a human fecal sample. The method and apparatus may be used by healthcare providers to distinguish Crohn's disease from other gastrointestinal illnesses, such as ulcerative colitis and irritable bowel syndrome.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of priority to U.S. application Ser. No. 10 / 280,564 filed on Oct. 25, 2002 which claims the benefit of priority to U.S. Provisional Application No. 60 / 335,812 filed on Oct. 26, 2001, the entirety of the disclosures of which are hereby incorporated by reference.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT [0002] Not Applicable. FIELD OF THE INVENTION [0003] A method and apparatus for the differentiation of Crohn's disease from other gastrointestinal illnesses, such as ulcerative colitis and irritable bowel syndrome, using the presence of fecal anti-Saccharomyces cerevisiae antibodies (ASCA) as a marker for Crohn's disease are provided. The apparatus includes an enzyme-linked immunoassay or other immunoassay that utilizes antibodies specific to human immunoglobulins for the measurement of total endogenous ASCA in a human fecal sample. The method and apparatus may be used by healthcare...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/564G01N33/569
CPCG01N33/564Y10S435/975G01N2469/20
Inventor BOONE, JAMES HUNTERLYERLY, DAVID MAXWELLWILKINS, TRACY DALE
Owner TECH LAB
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