Diagnosis Method Of Alcoholic Or Non-Alcoholic Steato-Hepatitis Using Biochemical Markers

Abstract

The present invention is drawn to a new diagnosis method for detecting the extent of alcoholic or non-alcoholic steato-hepatitis in a patient, in particular in a patient suffering from a disease involving alcoholic or non-alcoholic steato-hepatitis or who already had a positive diagnosis test of liver fibrosis and/or presence of liver necroinfiammatory lesions, by using the serum concentration of easily detectable biological markers. The invention is also drawn to diagnosis kits for the implementation of the method.

1 . An in vitro method for diagnosis of alcoholic or non-alcoholic steato-hepatitis or from a serum or plasma sample of a patient, comprising the steps of:
 a) studying 3 biochemical markers by measuring the values of their concentration in the serum or plasma of said patient, wherein said markers are:
 ApoA1 (apolipoprotein A1), 
 ALT (alanine aminotransferase), 
 AST (aspartate aminotransferase). 
   b) combining said values through a logistic function including said markers in order to obtain an end value, wherein said logistic function is obtained through the following method:   i) classification of a cohort of patients in different groups according to the extent of their disease;   ii) identification of factors which differ significantly between these groups by unidimensional analysis;   iii) logistic regression analysis to assess the independent discriminative value of markers for the diagnosis of alcoholic or non-alcoholic steato-hepatitis;   iv) construction of the logistic function by combination of these identified independent factors.   
     
     
         2 . The method of  claim 1 , wherein said logistic function further takes the age and gender of the patient into account. 
     
     
         3 . The method of  claim 2 , wherein said logistic function further comprises at least one biochemical marker chosen in the group constituted of alpha.2-macroglobulin, GGT (gammaglutamyl transpeptidase), total bilirubin, and haptoglobin. 
     
     
         4 . The method of  claim 3 , wherein said logistic function comprises all listed biochemical markers. 
     
     
         5 . The method of  claim 4 , wherein said logistic function is:
 f=a1−a2.[Age(years)]+a3.[ApoA1 (g/L)]−a4.Log[.alpha.2-macroglobulin (g/L)]+a5.Log[ALT (alanine aminotransferase)(IU/L)]−a6.Log[AST (aspartate aminotransferase)(IU/L )]+a7.Log[Total bilirubin (μmol/l)]−a8.Log[GGT (gammaglutamyl transpeptidase)(IU/L)]−a9.Log[Haptoglobin (g/L)]+a10.[Gender (female=0, male=1)], with   a1 comprised in the interval of [1.38435−90%; 1.38435+90%],   a2 comprised in the interval of [2.39829E−02−90%; 2.39829E−02+90%],   a3 comprised in the interval of [4.07571−90%; 4.07571+90%],   a4 comprised in the interval of [1.08306−90%; 1.08306+90%],   a5 comprised in the interval of [3.97299−90%; 3.97299+90%],   a6 comprised in the interval of [4.51309−90%; 4.51309+90%],   a7 comprised in the interval of [0.24014−90%; 0.24014+90%],   a8 comprised in the interval of [0.85462−90%; 0.85462+90%],   a9 comprised in the interval of [0.44638−90%; 0.44638+90%], and   a10 comprised in the interval of [0.86471−90%; 0.86471+90%].   
     
     
         6 . The method of  claim 5 , wherein said logistic function is:
 f=1.38435−2.39829E−02.[Age (years)]+4.07571.[ApoA1 (g/L)]−1.08306.Log[.alpha.2-macroglobulin (g/L)]+3.97299.Log[ALT (alanine aminotransferase)(IU/L)]−4.51309.Log[AST (aspartate aminotransferase)(IU/L)]+0.24014.Log[Total bilirubin (μmol/l)]−0.85462.Log[GGT (gammaglutamyl transpeptidase)(IU/L)]−0.446383.Log[Haptoglobin (g/L)]+0.86471.[Gender (female=0, male=1)].   
     
     
         7 . The method of  claim 1 , wherein said patient suffers from a disease involving alcoholic or non-alcoholic steato-hepatitis. 
     
     
         8 . The method of  claim 7 , wherein said disease is included in the group consisting of hepatitis B and C, alcoholism, hemochromatosis, metabolic disease, diabetes, obesity, autoimmune liver disease, primary biliary cirrhosis, alpha.1-antitrypsin deficit and Wilson disease. 
     
     
         9 . The method of  claims 1 , wherein said patient was already subjected to a diagnosis test of liver fibrosis and/or presence of liver necroinflammatory lesions. 
     
     
         10 . The method of  claim 9 , wherein said diagnosis test was FibroTest/Acti-Test. 
     
     
         11 . A kit for diagnosis of alcoholic or non-alcoholic steato-hepatitis in a patient comprising:
 a) instructions for determining the presence or absence of alcoholic or non-alcoholic steato-hepatitis in said patient;   b) reagents for measuring the serum values of the concentrations of 3 biochemical markers, wherein said markers are:
 ApoA1 (apolipoprotein A1), 
 ALT (alanine aminotransferase), and 
 AST (aspartate aminotransferase). 
   c) optionally, at least one reagent for measuring the serum values of the concentrations of alpha.2-macro globulin, GGT (gammaglutamyl transpeptidase), total bilirubin, and/or haptoglobin.   d) instructions for using a logistic function that is used to combine said values in order to obtain an end value, wherein analysis of said end value determines the presence or absence of alcoholic or non-alcoholic steato-hepatitis in said patient.   
     
     
         12 . The kit of  claim 11 , further comprising an ROC curve. 
     
     
         13 . The kit of  claim 11 , further comprising tables that allow to obtain the predictive values, depending on the expected prevalence of alcoholic or non-alcoholic steato-hepatitis in the patient population. 
     
     
         14 . The kit of  claim 11 , further comprising instructions for the quantification of alcoholic or non-alcoholic steato-hepatitis different grades (none, mild, moderate, or severe), and other intermediate grades.