Kits for detecting cervical dysplasia

Abstract

This invention provides methods and kits for improved diagnosis of medically relevant conditions by solution based biochemical testing procedures performed in solutions of test samples. The invention provides a method to substitute the cell based morphological information contained within the cytological and/or histological data of the test sample by molecular information obtainable from the solution, wherein the original test sample is dissolved and thus enables for accurate and reproducible assessment of medically relevant diagnosis from dissolved test samples. The method according to the invention comprises the steps of determining the levels of one or more disease markers associated with the condition to be diagnosed, determining the level of one or more normalization markers suitable to substitute the information related to morphological aspects of the sample, comparing and/or combining the data of the disease and normalization markers, and assessing diagnosis of a medically relevant condition.

1 . A test kit for detecting cervical dysplasia, comprising:
 (a) a first reagent for detecting a first marker molecule characteristic for cervical dysplasias selected from the group consisting of p16 INK4a , p14ARF, cyclin E, cyclin A, cyclin B, MN, her2/neu, mdm-2, bcl-2, EGF-Receptor, mcm-2, mcm-5, claudin-1, markers indicative for human papilloma virus infection, pRb, and p53; and   (b) a second reagent for detecting a normalization marker molecular characteristic for the presence or absence of epithelial cells selected from the group consisting of CK8, Ep-Cam, CK13, CK8, CK8, E-Cadherin, alpha-Catenin, beta-Catenin, gamma-Catenin, and involucrin.   
     
     
         2 . The test kit according to  claim 1 , wherein at least one of the first reagent and the second reagent is fixed to a solid phase. 
     
     
         3 . The test kit according to  claim 1 , furthermore comprising at least one of the following components:
 (a) the first marker molecular,   (b) the normalization marker molecular, and   (c) reagents and buffers commonly used for carrying out the detection reaction.   
     
     
         4 . The test kit according to  claim 1 , wherein the reagents for detecting the first marker molecule and the normal marker molecule comprise binding agents specific for said marker molecules. 
     
     
         5 . The test kit according to  claim 4 , wherein the binding agents are an antibody, a miniantibody, or a peptidomimetic comprising an antigen binding epitope. 
     
     
         6 . The test kit according to  claim 1 , wherein the test kit is a diagnostic test kit, an in-vitro diagnostic device, a research kit, or an analytical kit. 
     
     
         7 . The test kit according to  claim 1 , wherein the first marker molecule is p16 INK4a , and the normalization marker molecule is gamma-Catenin. 
     
     
         8 . The test kit according to  claim 7 , further comprising a reagent for the detection of Ep-Cam. 
     
     
         9 . The test kit according to  claim 7 , further comprising a buffer for sample lysis. 
     
     
         10 . The test kit according to  claim 7 , wherein the reagents for detecting p16 INK4a  and gamma-Catenin are fixed to solid phases. 
     
     
         11 . The test kit according to  claim 7 , wherein the test kit is an in-vitro diagnostic device.