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) measuring a level of sTNFRII in a sample obtained from a patient suspected of having an inflammatory bowel disease; and    b) 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 2  further comprising 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 . The method of  claim 1  further comprising acts of: 
 i) measuring in said sample or another sample obtained from said patient one or more additional cytokine level selected from the group consisting of IL-1β, IL-12p70, IL-10, IL-2, GM-CSF, TNF, IL-8, IL-4, IL-5, IL-6, Eotaxin, IFN-α, IFN-γ, sIL-6R, IL-12(total), IL-13, MIP-1β, MCP-1 and RANTES; and    ii) diagnosing from said sTNFRII level and said one or more additional cytokine levels the presence or absence in said patient of inflammatory bowel disease.    
   
   
       7 . The method of  claim 6 , wherein said diagnosing step comprises comparing said sTNFRII level and said one or more additional cytokine levels 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 one or more additional cytokine levels.  
   
   
       9 . The method of  claim 8 , wherein said distinguishing ulcerative colitis from Crohn's disease comprises comparing said sTNFRII level and said one or more additional cytokine levels to a cytokine profile indicative of Crohn's disease or ulcerative colitis.  
   
   
       10 . The method as in  claim 7 , wherein said sTNFRII level above a sTNFRII detection cut-off value and said additional one or more cytokine levels below a cytokine detection cut-off value are considered indicative of inflammatory bowel disease.  
   
   
       11 . The method as in  claim 7 , wherein a ratio of said sTNFRII level to an additional cytokine level above a detection cut-off ratio value is considered indicative of inflammatory bowel disease.  
   
   
       12 . The method as in  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) measuring the level of a first analyte in a sample obtained from a patient suspected of having inflammatory bowel disease;    b) measuring in said sample or another sample obtained from said patient the level of one or more additional analytes, wherein said one or more additional analytes are different form said first analyte; and    c) diagnosing from said first analyte level and said one or more additional analyte levels the presence or absence in said patient of inflammatory bowel disease.    
   
   
       18 . The method of  claim 17 , wherein said first analyte is a cytokine and wherein said one or more additional analytes comprise one or more additional cytokines.  
   
   
       19 . The method of  claim 18 , wherein said first cytokine is selected from a group consisting of IL-1β, IL-12p70, IL-10, IL-2, GM-CSF, TNF, IL-8, IL-4, IL-5, IL-6, Eotaxin, IFN-α, IFN-γ, sIL-6R, IL-12(total), IL-13, MIP-1 P, MCP-1, RANTES and sTNFRII.  
   
   
       20 . The method of  claim 18 , wherein said one or more additional cytokines is selected from the group consisting of IL-1β, IL-12p70, IL-10, IL-2, GM-CSF, TNF, IL-8, IL-4, IL-5, IL-6, Eotaxin, IFN-a, IFN-γ, sIL-6R, IL-12(total), IL-13, MIP-1β, MCP-1, RANTES and sTNFRII.  
   
   
       21 . The method of  claim 18 , wherein said determining step comprises comparing said first cytokine level and said one or more additional cytokine levels to a cytokine profile indicative of inflammatory bowel disease.  
   
   
       22 . The method of  claim 18  further comprising distinguishing ulcerative colitis from Crohn's disease on the basis of said first cytokine level and said one or more additional cytokine levels.  
   
   
       23 . The method of  claim 22 , wherein said distinguishing ulcerative colitis from Crohn's disease comprises comparing said first cytokine level and said one or more additional cytokine levels to profiles indicative of Crohn's disease or ulcerative colitis.  
   
   
       24 . The method of  claim 18 , wherein a ratio of said first cytokine level to said additional cytokine level above a detection cut-off ratio value is considered indicative of inflammatory bowel disease.  
   
   
       25 . The method of  claim 22  further comprising distinguishing ulcerative colitis from Crohn's disease by comparing said first cytokine level to a first cytokine discrimination cut-off value, wherein first cytokine 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.  
   
   
       26 . The method of  claim 22  further comprising distinguishing ulcerative colitis from Crohn's disease by comparing said first cytokine level to a first cytokine discrimination cut-off value, wherein first cytokine 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.  
   
   
       27 . The method of  claim 22  further comprising distinguishing ulcerative colitis from Crohn's disease by comparing said cytokine levels to a cytokine 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.  
   
   
       28 . The method of  claim 1 , wherein said measuring is conducted on a single sample.  
   
   
       29 . The method of  claim 1 , wherein said measuring is conducted in a single assay chamber.  
   
   
       30 . The method of  claim 1 , wherein said assay chamber is a single well of an assay plate.  
   
   
       31 . The method of  claim 1 , wherein said assay chamber is an assay chamber of a cartridge.  
   
   
       32 . The method of  claim 1  further comprising conducting a diagnostics test to determine if said patient has viral or bacterial infection.  
   
   
       33 . The method of  claim 1 , wherein said sample is blood, serum or plasma.  
   
   
       34 . The method of  claim 1 , wherein said sample is a fecal sample.  
   
   
       35 . The method of  claim 1 , wherein said sample is selected from a group consisting of biopsy tissue, intestinal mucosa or urine.  
   
   
       36 . The method of  claim 1 , wherein said level is measured using an immunoassay.  
   
   
       37 . The method of  claim 1  further comprising determining from said level of sTNFRII the extent of inflammation from said disease.  
   
   
       38 . The method of  claim 17  further comprising determining from said measured levels the extent of inflammation from said disease.  
   
   
       39 . A method for monitoring the progression or treatment of an inflammatory bowel disease comprising acts of: 
 (a) measuring the levels of sTNFRII in samples obtained, at different times, from a patient that has or is suspected to have an inflammatory bowel disease; and    (b) determining from said levels of sTNFRII the progression or efficacy of treatment of the disease.    
   
   
       40 . A method for monitoring the progression or treatment of an inflammatory bowel disease comprising acts of: 
 a) measuring the levels of a first analyte in samples obtained, at different times, from a patient that has or is suspected to have an inflammatory bowel disease;    b) measuring the levels of one or more additional analytes in said samples, wherein said one or more additional analytes differ from said first analyte; and    c) determining from said levels the progression or efficacy of treatment of the disease.    
   
   
       41 . The method of  claim 40 , wherein said first analyte is a cytokine and wherein said one or more additional analytes comprise one or more additional cytokines.  
   
   
       42 . The method as in  claim 41 , wherein said first cytokine is selected from a group consisting of IL-1β, IL-12p70, IL-10, IL-2, GM-CSF, TNF, IL-8, IL-4, IL-5, IL-6, Eotaxin, IFN-α, IFN-γ, sIL-6R, IL-12(total), IL-13, MIP-1β, MCP-1, RANTES and sTNFRII.  
   
   
       43 . The method as in  claim 41 , wherein said one or more additional cytokines are selected from the group consisting of IL-1β, IL-12p70, IL-10, IL-2, GM-CSF, TNF, IL-8, IL-4, IL-5, IL-6, Eotaxin, IFN-α, IFN-γ, sIL-6R, IL-12(total), IL-13, MIP-1β, MCP-1, RANTES and sTNFRII.  
   
   
       44 . A method for evaluation of the effectiveness of a drug and/or drug candidate in treating an inflammatory bowel disease comprising acts of: 
 (a) measuring the level of 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; and    (b) determining from said level the effectiveness of the drug or drug candidate.    
   
   
       45 . A method for evaluation of the effectiveness of a drug and/or drug candidate in treating an inflammatory bowel disease comprising acts of: 
 (a) measuring the level of a first analyte 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) measuring 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 the level of one or more additional analytes, wherein one or more additional analytes differ from said first analyte; and    c) determining from said levels the effectiveness of the drug or drug candidate.    
   
   
       46 . The method of  claim 45 , wherein said first analyte is a cytokine and wherein said one or more additional analytes comprise one or more additional cytokines.  
   
   
       47 . The method of  claim 46 , wherein said first cytokine is selected from a group consisting of IL-1β, IL-12p70, IL-10, IL-2, GM-CSF, TNF, IL-8, IL-4, IL-5, IL-6, Eotaxin, IFN-α, IFN-γ, sIL-6R, IL-12(total), IL-13, MIP-1β, MCP-1, RANTES and sTNFRII.  
   
   
       48 . The method of  claim 46 , wherein said one or more additional cytokines are selected from the group consisting of IL-1β, IL-12p70, IL-10, IL-2, GM-CSF, TNF, IL-8, IL-4, IL-5, IL-6, Eotaxin, IFN-α, IFN-γ, sIL-6R, IL-12(total), IL-13, MIP-1β, MCP-1, RANTES and sTNFRII.  
   
   
       49 . A method for diagnosing an inflammatory bowel disease (IBD) comprising acts of: 
 a) measuring a level of a cytokine in a sample obtained by a non-surgically invasive procedure from a human patient suspected of having an inflammatory bowel disease; and    b) diagnosing from said level the presence or absence in said patient of inflammatory bowel disease.    
   
   
       50 . The method of  claim 49 , wherein said cytokine is selected from a group consisting of IL-1β, IL-12p70, IL-10, IL-2, GM-CSF, TNF, IL-8, IL-4, IL-5, IL-6, Eotaxin, IFN-α, IFN-γ, sIL-6R, IL-12(total), IL-13, MIP-1β, MCP-1, RANTES and sTNFRII.  
   
   
       51 . The method of  claim 49 , wherein said sample is blood, serum, plasma, a fecal sample, or urine.  
   
   
       52 . The method of  claim 18 , wherein said first cytokine is sTNFRII and said one or more additional cytokine is selected from a group consisting of RANTES, IL-6R and IL-4.