Diagnostic test

Abstract

Disclosed are methods for conducting diagnostic tests for the detection of the inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis. Also described are methods for monitoring a patient by administering tests of the present invention. Also described are methods for monitoring patient's treatment by administering tests of the present invention. Also described are methods for evaluating the effectiveness of a drug or a drug candidate by administering tests of the present invention to samples from patients, animal models, and cell cultures treated with a drug or a drug candidate. Also disclosed are methods for determining the usefulness of analytes, e.g. cytokines, for acting as diagnostic and monitoring markers for inflammatory bowel disease in the various methods of the invention.

1 . A method for diagnosing an inflammatory bowel disease (IBD) comprising acts of:
 (a) obtaining a sample from a patient suspected of having an inflammatory bowel disease;   (b) performing an assay to measure a level of soluble TNF receptor II (sTNFRII) in the sample obtained from a patient suspected of having an inflammatory bowel disease; and   (c) diagnosing from said level the presence or absence in said patient of inflammatory bowel disease.   
     
     
         2 . The method of  claim 1 , wherein said inflammatory bowel disease is ulcerative colitis. 
     
     
         3 . The method of  claim 1 , wherein said diagnosing step comprises comparing said measured level to a detection cut-off value, wherein said sTNFRII level above said detection cut-off value is considered indicative of inflammatory bowel disease. 
     
     
         4 . The method of  claim 1 , wherein said diagnosing step further comprises distinguishing ulcerative colitis from Crohn's disease on the basis of said level of sTNFRII. 
     
     
         5 . The method of  claim 4 , wherein said distinguishing ulcerative colitis from Crohn's disease comprises comparing said measured level to a discrimination cut-off value, wherein said level below said discrimination cut-off value is considered indicative of Crohn's disease and above said discrimination cut-off value is considered indicative of ulcerative colitis. 
     
     
         6 . A method for diagnosing an inflammatory bowel disease (IBD) comprising acts of:
 (a) obtaining a sample from a patient suspected of having an inflammatory bowel disease;   (b) performing an assay to measure a level of soluble TNF receptor II (sTNFRII) in said sample;   (c) performing an assay to measure in said sample or another sample obtained from said patient a level of MIP-1β; and   (d) diagnosing from said sTNFRII level and said level of MIP-1β the presence or absence in said patient of inflammatory bowel disease.   
     
     
         7 . The method of  claim 6 , wherein said diagnosing step further comprises comparing said sTNFRII level and said level of MIP-1β to a cytokine profile indicative of inflammatory bowel disease. 
     
     
         8 . The method of  claim 6  further comprising distinguishing ulcerative colitis from Crohn's disease on the basis of said sTNFRII level and said level of MIP-1β. 
     
     
         9 . The method of  claim 8 , wherein said distinguishing ulcerative colitis from Crohn's disease comprises comparing said sTNFRII level and said level of MIP-1β to a cytokine profile indicative of Crohn's disease or ulcerative colitis. 
     
     
         10 . The method of  claim 7 , wherein said sTNFRII level above a sTNFRII detection cut-off value and said level of MIP-1βbelow a cytokine detection cut-off value are considered indicative of inflammatory bowel disease. 
     
     
         11 . The method of  claim 7 , wherein a ratio of said sTNFRII level to said MIP-1βlevel above a detection cut-off ratio value is considered indicative of inflammatory bowel disease. 
     
     
         12 . The method of  claim 7 , wherein said sTNFRII level being above a sTNFRII detection cut-off line, curve, or surface on a correlation plot is considered indicative in inflammatory bowel disease. 
     
     
         13 . The method of  claim 9  further comprising distinguishing ulcerative colitis from Crohn's disease by comparing said sTNFRII level to a sTNFRII discrimination cut-off value, wherein sTNFRII levels below said sTNFRII discrimination cut-off value are considered indicative of Crohn's disease and above said sTNFRII discrimination cut-off value are considered indicative of ulcerative colitis. 
     
     
         14 . The method of  claim 9  further comprising distinguishing ulcerative colitis from Crohn's disease by comparing said sTNFRII level to a sTNFRII discrimination cut-off line, curve, or surface on a correlation plot, wherein sTNFRII levels below said sTNFRII discrimination cut-off line, curve, or surface are considered indicative of Crohn's disease and above said sTNFRII discrimination cut-off line, curve, or surface are considered indicative of ulcerative colitis. 
     
     
         15 . The method of  claim 3 , wherein said sTNFRII detection cut-off level is between 5 and 7 ng per ml of sample. 
     
     
         16 . The method of  claim 5 , wherein said sTNFRII discrimination cut-off level is between 8 and 10 ng per ml of sample. 
     
     
         17 . A method for diagnosing inflammatory bowel disease comprising acts of:
 a) obtaining a sample from a patient suspected of having an inflammatory bowel disease and performing an assay to measure a level of soluble TNF receptor II (sTNFRII) in said sample;   b) performing an assay to measure in said sample or another sample obtained from said patient a level of an additional analyte, wherein said additional analyte is a cytokine; and   c) diagnosing from said sTNFRII level and said additional analyte level the presence or absence in said patient of inflammatory bowel disease.   
     
     
         18 . The method of  claim 17 , wherein said additional analyte is MIP-1β. 
     
     
         19 . The method of  claim 17 , wherein said diagnosing step comprises comparing said sTNFRII level and said additional analyte level to an analyte profile indicative of inflammatory bowel disease. 
     
     
         20 . The method of  claim 17 , wherein said diagnosing step further comprises distinguishing ulcerative colitis from Crohn's disease on the basis of said sTNFRII level and said additional analyte level. 
     
     
         21 . The method of  claim 20 , wherein said distinguishing ulcerative colitis from Crohn's disease comprises comparing said analyte level and said additional analyte level to profiles indicative of Crohn's disease or ulcerative colitis. 
     
     
         22 . The method of  claim 17 , wherein a ratio of said sTNFRII level to said additional analyte level above a detection cut-off ratio value is considered indicative of inflammatory bowel disease. 
     
     
         23 . The method of  claim 20  wherein said distinguishing ulcerative colitis from Crohn's disease comprises comparing said sTNFRII level to a sTNFRII discrimination cut-off value, wherein sTNFRII levels above said discrimination cut-off value are considered indicative of Crohn's disease and below said discrimination cut-off value are considered indicative of ulcerative colitis. 
     
     
         24 . The method of  claim 20  wherein said distinguishing ulcerative colitis from Crohn's disease comprises comparing said sTNFRII level to a sTNFRII discrimination cut-off value, wherein sTNFRII levels below said discrimination cut-off value are considered indicative of Crohn's and above said discrimination cut-off value are considered indicative of ulcerative colitis. 
     
     
         25 . The method of  claim 20  wherein said distinguishing ulcerative colitis from Crohn's disease comprises comparing said sTNFRII levels to a sTNFRII profile defined as at least one area or volume situated between a first detection cut-off line, curve, or surface and a second discrimination cut-off line, curve, or surface on a correlation plot. 
     
     
         26 . The method of  claim 1 , wherein said measuring step is conducted on a single sample. 
     
     
         27 . The method of  claim 1 , wherein said measuring step is conducted in a single assay chamber. 
     
     
         28 . The method of  claim 27 , wherein said assay chamber is a single well of an assay plate. 
     
     
         29 . The method of  claim 27 , wherein said assay chamber is an assay chamber of a cartridge. 
     
     
         30 . The method of  claim 1  further comprising conducting a diagnostic test to determine if said patient has viral or bacterial infection. 
     
     
         31 . The method of  claim 1 , wherein said sample is selected from the group consisting of blood, serum or plasma. 
     
     
         32 . The method of  claim 1 , wherein said sample is a fecal sample. 
     
     
         33 . The method of  claim 1 , wherein said sample is selected from the group consisting of biopsy tissue, intestinal mucosa or urine. 
     
     
         34 . The method of  claim 1 , wherein said level is measured using an immunoassay. 
     
     
         35 . The method of  claim 1  further comprising determining from said level of sTNFRII the extent of inflammation from said disease. 
     
     
         36 . The method of  claim 17  further comprising determining from said measured levels the extent of inflammation from said disease. 
     
     
         37 . A method for monitoring the progression or treatment of an inflammatory bowel disease comprising acts of:
 (a) obtaining a plurality of samples at different points in time from a patient that has or is suspected of having an inflammatory bowel disease and performing an assay to measure levels of soluble TNF receptor II (sTNFRII) in said samples; and   (b) determining from said levels of sTNFRII the progression or efficacy of treatment of the disease.   
     
     
         38 . A method for monitoring the progression or treatment of an inflammatory bowel disease comprising acts of:
 a) obtaining a plurality of samples at different points in time from a patient that has or is suspected of having an inflammatory bowel disease and performing an assay to measure levels of soluble TNF receptor II (sTNFRII) in said samples;   b) performing an assay to measure levels of an additional analyte in said samples, wherein said additional analyte is a cytokine; and   c) determining from said levels the progression or efficacy of treatment of the disease.   
     
     
         39 . The method as in  claim 38 , wherein said additional analyte is MIP-1β. 
     
     
         40 . A method for evaluation of the effectiveness of a drug and/or drug candidate in treating an inflammatory bowel disease (IBD) comprising acts of:
 (a) performing an assay to measure a level of soluble TNF receptor II (sTNFRII) in a sample obtained from a human, non-human animal with IBD or a model system of IBD exposed to a drug or drug candidate;   (b) performing an assay to measure a level of sTNFRII in a control sample obtained from a human or non-human animal that was not exposed to the drug or drug candidate;   (c) comparing the sTNFRII levels measured in steps (a) and (b); and   (d) determining from said comparing step (c) the effectiveness of the drug or drug candidate.   
     
     
         41 . A method for evaluation of the effectiveness of a drug and/or drug candidate in treating an inflammatory bowel disease (IBD) comprising acts of:
 (a) performing an assay to measure a level of soluble TNF receptor II (sTNFRII) in a sample obtained from a human, non-human animal with IBD or a model system of IBD exposed to a drug or drug candidate;   (b) performing an assay to measure in said sample or another sample obtained from said human, non-human animal with IBD or a model system of IBD exposed to said drug or drug candidate a level of an additional analyte, wherein said additional analyte is a cytokine;   (c) performing an assay to measure a level of sTNFRII in a control sample obtained from a human or non-human animal that was not exposed to the drug or drug candidate;   (d) performing an assay to measure a level of said additional analyte in said control sample or another control sample obtained from a human or non-human animal that was not exposed to the drug or drug candidate;   (e) comparing the sTNFRII levels measured in steps (a) and (c) and comparing the levels of said additional analyte measured in steps (b) and (d); and   (f) determining from said comparing step (e) the effectiveness of the drug or drug candidate.   
     
     
         42 . The method of  claim 41 , wherein said additional analyte is MIP-1β. 
     
     
         43 . A method for diagnosing an inflammatory bowel disease (IBD) comprising acts of:
 (a) obtaining a sample by a non-surgically invasive procedure from a human patient suspected of having an inflammatory bowel disease;   (b) performing an assay to measure a level of a sTNFRII in said sample; and   (c) diagnosing from said level the presence or absence in said patient of inflammatory bowel disease.   
     
     
         44 . The method of  claim 43 , wherein said sample is selected from the group consisting of blood, serum, plasma, a fecal sample, or urine. 
     
     
         45 . The method of  claim 17 , wherein said additional analyte is selected from a group consisting of RANTES, IL-6R and IL-4. 
     
     
         46 . The method of  claim 1 , wherein said assay is selected from the group consisting of: an immunochromatographic assay; a solid-phase assay and an agglutination assay. 
     
     
         47 . The method of  claim 6 , wherein said assay to measure a level of sTNFRII and said assay to measure a level of MIP-1β are selected from the group consisting of: an immunoassay; an immunochromatographic assay; a solid-phase assay and an agglutination assay. 
     
     
         48 . The method of  claim 17 , wherein said assay to measure a level of sTNFRII and said assay to measure a level of an additional analyte are selected from the group consisting of: an immunoassay; an immunochromatographic assay; a solid-phase assay and an agglutination assay. 
     
     
         49 . The method of  claim 37 , wherein said assay is selected from the group consisting of: an immunoassay; an immunochromatographic assay; a solid-phase assay and an agglutination assay. 
     
     
         50 . The method of  claim 38 , wherein said assay to measure a level of sTNFRII and said assay to measure a level of an additional analyte are selected from the group consisting of: an immunoassay; an immunochromatographic assay; a solid-phase assay and an agglutination assay. 
     
     
         51 . The method of  claim 40 , wherein said assay to measure a level of sTNFRII is selected from the group consisting of: an immunoassay; an immunochromatographic assay; a solid-phase assay and an agglutination assay. 
     
     
         52 . The method of  claim 41 , wherein said assay to measure a level of sTNFRII and said assay to measure a level of an additional analyte are selected from the group consisting of: an immunoassay; an immunochromatographic assay; a solid-phase assay and an agglutination assay.