METHOD FOR DETECTING HUMAN PAPILLOMAVIRUS mRNA

Abstract

An in vitro method is provided for screening human female subjects to assess their risk of developing cervical carcinoma which comprises screening the subject for expression of mRNA transcripts from the E6 and optionally the L1 gene of human papillomavirus, wherein subjects positive for expression of L1 and/or E6 mRNA are scored as being at risk of developing cervical carcinoma. Kits for carrying out such methods are also provided.

1 . An in vitro method of screening human subjects to assess their risk of developing cervical carcinoma, which method comprises screening the subject for expression of mRNA transcripts of the E6 gene of HPV and sorting the subject into one of two categories of risk for development of cervical carcinoma based on expression of E6 mRNA, wherein individuals positive for expression of E6 mRNA are scored as carrying integrated HPV or a modified episomal HPV genome and are therefore classified as high risk for development of cervical carcinoma, whereas individuals negative for expression of E6 mRNA are scored as not carrying integrated HPV or a modified episomal HPV genome and are therefore classified as no detectable risk for development of cervical carcinoma. 
     
     
         2 . An in vitro method of identifying human subjects having abnormal cell changes in the cervix, which method comprises screening the subject for expression of mRNA transcripts of the E6 gene of HPV, wherein individuals positive for expression of E6 mRNA are identified as having abnormal cell changes in the cervix. 
     
     
         3 . A method according to  claim 1  wherein the human subjects are subjects previously identified as infected with human papillomavirus DNA in cells of the cervix. 
     
     
         4 . A method according to  claim 1  wherein the human subjects are subjects having a previous diagnosis ASCUS, CIN 1 lesions or condyloma. 
     
     
         5 . A method according to  claim 1  wherein individuals positive for expression of E6 mRNA from at least one of HPV types 16, 18, 31, 33 or 45 are scored as carrying integrated HPV. 
     
     
         6 . A method according to  claim 1  which comprises screening for E6 mRNA expression using a technique which is able to detect E6 mRNA from at least one cancer-associated HPV type, 
     
     
         7 . A method according to  claim 1  which comprises screening for E6 mRNA expression using a technique which is able to detect E6 mRNA from HPV types 16, 18, 31, 33, and 45. 
     
     
         8 . A method according to  claim 1  wherein screening for E6 mRNA expression is carried using an amplification reaction to amplify of a region of the mRNA, together with real-time detection of the products of the amplification reaction. 
     
     
         9 . A method according to  claim 1  wherein screening for E6 mRNA expression is carried using real-time NASBA. 
     
     
         10 . A kit for use in the detection of mRNA transcripts of the E6 genes of HPV, the kit comprising primer-pairs which enable amplification of a region of E6 transcripts from HPV types 16, 18, 31, 33, and 45.