Novel diagnostic marker, a diagnostic kit and a method for diagnosis of rheumatoid arthritis

Abstract

The present invention relates to a novel diagnostic marker useful for the diagnosis of rheumatoid arthritis comprising the autoantibodies of mannose binding lectin protein and a process thereof.

1 . Novel diagnostic marker useful for the diagnosis of rheumatoid arthritis comprising the autoantibodies of mannose binding lectin (MBL) protein.  
   
   
       2 . A novel diagnostic marker as claimed in  claim 1 , wherein the presence of said marker indicates the possibility of rheumatoid arthritis.  
   
   
       3 . A novel diagnostic marker as claimed in  claim 1 , wherein the specific level of said marker relates with the level of disease activity and the level of joint damage.  
   
   
       4 . A method for the diagnosis of rheumatoid arthritis in the human subject, wherein the said method comprising the steps of: 
 a) Screening of normal control individuals and rheumatoid arthritis patients for the diagnostic marker as claimed in  claim 1  by quantifying the level of autoantibodies to MBL by ELISA in the blood of a human subject.    b) Statistically comparing the levels of autoantibodies to MBL in the rhematoid arthritis (RA) patients with that of the normal healthy individuals obtained from step (a) to establish association of rheumatoid arthritis disease with the elevated MBL auto antibodies.    c) Estimating the isotypes of rheumatoid factors (IgM RF and IgG RF) by the ELISA test followed by computing their association with rheumatoid arthritis disease.    d) Comparing the values of auto antibodies to MBL and the RF isotypes (IgM RF and IgG RF) obtained from step (c) for the diagnosis of RA, wherein the presence of anti MBL autoantibodies indicates the possibility of the rhematoid arthritis.    e) A method as claimed in  claim 4 , wherein the blood of normal control individuals are collected from Guru Teg Bahadur Hospital, Delhi, India.    
   
   
       5 . A method as claimed in  claim 4 , wherein the blood of RA patients are collected from Army Hospital, Department of Rheumatology, New Delhi, India.  
   
   
       6 . A method as claimed in  claim 4 , wherein the level of MBL auto antibodies in a RA affected patient is significantly higher than a normal healthy individual.  
   
   
       7 . A method as claimed in  claim 4 , wherein the levels of MBL auto antibodies in RA patients is in the range from 311 to 1192 AU/ml.  
   
   
       8 . A method as claimed in  claim 4 , wherein the levels of MBL auto antibodies in the healthy controls is in the range from 177 to 752 AU/ml.  
   
   
       9 . A method as claimed in  claim 4 , wherein the presence of significantly high levels of auto antibodies to MBL (P value<0.0001), is associated with high disease (RA) activity and joint damage in the RA patient as compared to healthy individuals with out any symptom of RA.  
   
   
       10 . A method as claimed in  claim 4 , wherein the computational analysis for the comparison of the autoantibodies to MBL and RF isotypes (IgM RF and IgG RF) are carried out by known statistical tools.  
   
   
       11 . A diagnostic kit for diagnosis of Rheumatoid arthritis comprising: 
 a purified or recombinant Mannose binding protein;    b anti-MBL auto antibody serum standard;    c requisite reconstituable coating, washing and dilution buffers;    d micro titre plates;    e secondary enzyme conjugated antibodies;    f appropriate substrate reagent, stop solution.