Methods and Devices for Diagnosis of Appendicitis

Abstract

A method is provided for diagnosing appendicitis in a patient that includes identifying at least one symptom of appendicitis in the patient and identifying the presence of at least one molecule differentially associated with appendicitis in a fluid or tissue sample of said patient. MRP-8/14 and haptoglobin are examples of molecules differentially associated with appendicitis. Devices and kits for performing the appendicitis assays of this invention are also provided. In one embodiment, the device is in a flow-through immunoassay format for testing blood samples. Further, methods for screening for molecules differentially associated with appendicitis are provided that include the use of samples from patients being operated on for suspected appendicitis.

1 . A method for identifying a molecule differentially associated with appendicitis, said method comprising:
 a) obtaining a sample of blood, serum or plasma from each of a plurality of patients who are undergoing surgery for suspected appendicitis;   b) determining during surgery whether each said patient has appendicitis or not; and   c) analyzing said samples for the presence of a molecule differentially associated with appendicitis, thereby identifying a molecule differentially associated with appendicitis.   
     
     
         2 . The method of  claim 1  wherein said analyzing is done by identifying a molecule that is present in the samples from patients who have appendicitis but not present in the samples from patients who do not have appendicitis. 
     
     
         3 . The method of  claim 1  wherein the amount of said molecule in the samples from patients who have appendicitis is determined and compared to the amount of said molecule in the samples of patients who do not have appendicitis to determine an amount of said molecule that is differentially associated with appendicitis. 
     
     
         4 . The method of  claim 1  wherein the samples are blood samples. 
     
     
         5 . The method of  claim 1  wherein the molecule differentially associated with appendicitis is selected from the group consisting of unique structural proteins of the gastrointestinal tract, stress-related inflammatory mediators, immunologic factors, indicators of intestinal bacterial flora, Plasminogen Activator Inhibitor-1, fatty acid binding proteins, nuclear factor kappa beta (NFκB), specific appendix antigens (HLA-DR), inflammation associated antigens, and nucleic acids coding for any of the foregoing. 
     
     
         6 . A method of detecting the presence of a molecule differentially associated with appendicitis in a sample from a patient suspected of having appendicitis comprising:
 a) obtaining a sample of blood, serum or plasma from a patient suspected of having appendicitis;   b) contacting said sample with an antibody to said molecule; and   c) reading the results of said means for detecting binding thereby detecting the presence of a molecule differentially associated with appendicitis in said sample.   
     
     
         7 . The method of  claim 6  wherein said sample is contacted with said antibody in an immunoassay device. 
     
     
         8 . The method of  claim 7  wherein said sample is serum. 
     
     
         9 . The method of  claim 7  wherein said sample is plasma. 
     
     
         10 . The method of  claim 7  wherein said sample is whole blood. 
     
     
         11 . The method of  claim 7  wherein said immunoassay device comprises:
 i) a first antibody specific to said molecule;   ii) a support for said first antibody;   iii) means for contacting said first antibody with said sample; and   iv) means for detecting binding of said first antibody with said molecule.   
     
     
         12 . A method for diagnosing appendicitis in a patient comprising:
 a) identifying at least one symptom of appendicitis in said patient; and   b) identifying the presence of at least one molecule differentially associated with appendicitis in a blood, serum or plasma sample of said patient;   
       thereby diagnosing appendicitis in said patient. 
     
     
         13 . The method of  claim 12  wherein said identification of said molecule is performed by detecting a nucleic acid sequence coding for said molecule in said sample. 
     
     
         14 . The method of  claim 13  wherein said molecule is MRP-8/14. 
     
     
         15 . The method of  claim 13  wherein said nucleic acid is detected in a blood sample. 
     
     
         16 . The method of  claim 13  wherein said nucleic acid is detected in a serum sample. 
     
     
         17 . The method of  claim 13  wherein said nucleic acid is detected in a plasma sample. 
     
     
         18 . An immunoassay test device for detecting the presence of a molecule differentially associated with appendicitis in a blood sample from a patient suspected of having appendicitis, said device comprising:
 a first antibody specific to MRP-8/14;   a support for said first antibody;   means for contacting said first antibody with said sample; and   means for detecting binding of said first antibody with said molecule.