Method for detecting carcinomas in a solubilized cervical body sample

Abstract

The present invention relates to a method for the early diagnosis of carcinomas and their preliminary stages, which comprises determining the overexpression of a cell cycle regulatory protein in a solubilized body sample. The present invention is particularly directed to a method for detecting cervical carcinomas, cervical intraepithelial neoplasias, or cervical carcinomas in-situ from a solubilized cervical body sample of a human subject, by solubilizing the cervical body sample in a lysis buffer, and determining the overexpression of cyclin dependent kinase inhibitor p16 in the solubilized cervical sample. The invention also concerns a test kit usable for this purpose as well as an in-vitro diagnostic device.

1 . An in-vitro diagnostic device comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed on solid carriers, for detecting p16 in a solubilized sample.  
   
   
       2 . The in-vitro diagnostic device according to  claim 1 , which is selected from the group consisting of: 
 a. an ELISA device comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to ELISA plates, ELISA stripes or ELISA wells;    b. a lateral flow test device, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to test strips, colloidal gold particles or latex particles;    c. a flow through assay device, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to a porous member, or to the surface of capillaries;    d. a latex agglutination assay device, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to latex particles;    e. an immunoassay device, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to beads or membranes; and    f. an immunoassay device, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to microspheres.    
   
   
       3 . A test kit for determining the level of cyclin dependent kinase inhibitor p16 comprising antibodies directed against cyclin dependent kinase inhibitor p16 and a lysis buffer for solubilization of a body sample.  
   
   
       4 . The test kit according to  claim 3 , wherein the lysis buffer comprises at least one composition selected from the group consisting of chaotropic agents, anionic detergents, cationic detergents, non-ionic detergents, amphoteric detergents, and alkaline compositions.  
   
   
       5 . The test kit according to  claim 3 , wherein the lysis buffer comprises at least one composition selected from the group consisting of a proteinase inhibitor, a DNAse inhibitor, and an RNAse inhibitor.  
   
   
       6 . The test kit according to  claim 5 , wherein the proteinase inhibitor is selected from the group consisting of inhibitors to serine proteinases, inhibitors to cysteine proteinases, inhibitors to aspartic proteinases, inhibitors to metallo proteinases, inhibitors to acidic proteinases, inhibitors to neutral proteinases, and inhibitors to alkaline proteinases.  
   
   
       7 . The test kit according to  claim 3 , wherein the lysis buffer comprises at least one non-ionic detergent and at least one proteinase inhibitor.  
   
   
       8 . The test kit according to  claim 7 , wherein the lysis buffer contains Triton X-100 and at least one inhibitor of serine proteineases.  
   
   
       9 . The test kit according to  claim 3 , further comprising at least one marker molecule for carrying out a positive control reactions, reagents and buffers commonly used for carrying out the detection reaction.  
   
   
       10 . The test kit according to  claim 3 , further comprising a recombinant p16 protein, fragments thereof or peptides derived from p16 as marker molecules for carrying out a positive control reaction.  
   
   
       11 . A test kit comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed on solid carriers, for detecting p16 in a solubilized sample.  
   
   
       12 . The test kit according to  claim 11 , which is selected from the group consisting of: 
 a. an ELISA kit comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to ELISA plates, ELISA stripes or ELISA wells;    b. a lateral flow test kit, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to test strips, colloidal gold particles or latex particles;    c. a flow through assay kit, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to a porous member, or to the surface of capillaries;    d. a latex agglutination assay kit, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to latex particles;    e. an immunoassay kit, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to beads or membranes; and    f. an immunoassay kit, comprising antibodies directed against cyclin dependent kinase inhibitor p16 fixed to microspheres.