Detection method for human pappilomavirus (HPV) and its application in cervical cancer

Abstract

Embodiments of the invention provide methods, assays, and kits for detecting HPV infection and HPV associated epithelial cell abnormalities, most notably those associated with pre-malignant and malignant epithelial cell lesions. Detection of HPV DNAs, genomes, and/or oncoproteins by nucleic acid hybridization assays and immunological assays can be used in early clinical screening for HPV infection and diagnosis for cervical cancer. The polypeptides, recombinant proteins, antibodies, nucleic acids, and various detection methods thereof are particularly useful for diagnosing carcinomas of the uterine cervix and those at risk of developing cervical cancer.

1 . A papillomavirus detection composition comprising a recombinant papillomavirus protein having native folding and being recognized by one or more papillomavirus antibodies present from a clinical sample. 
     
     
         2 . The papillomavirus detection composition of  claim 1 , wherein the clinical sample is selected from the group consisting of cervical cells, cervical tissues, cervical swabs, body fluids, serum, blood, tumors, and combination thereof. 
     
     
         3 . The papillomavirus detection composition of  claim 1 , wherein the recombinant papillomavirus protein is selected from the group consisting of recombinant HPV-16 E6 proteins, recombinant HPV-16 E7 proteins, recombinant HPV-18 E6 proteins, recombinant HPV-18 E7 proteins, recombinant HPV-16 L1 proteins, recombinant HPV-18 L1 proteins, and combinations thereof. 
     
     
         4 . The papillomavirus detection composition of  claim 1 , wherein the recombinant papillomavirus protein is purified as a soluble protein. 
     
     
         5 . The papillomavirus detection composition of  claim 1 , wherein the recognition of the recombinant papillomavirus protein to the one or more papillomavirus antibodies from the clinical sample is performed by conducting one or more immunological assays on the clinical sample. 
     
     
         6 . The papillomavirus detection composition of  claim 5 , wherein the one or more immunological assays are selected from the group consisting of ELISA assays, antigen assays for papillomavirus proteins, antibody assays for antibodies against papillomavirus proteins, assays for papillomavirus immunocomplexes, protein chip assays, radioimmunoprecipitation assays, rapid membrane immunochromatographic assays, rapid stick immunochromatographic assays. 
     
     
         7 . A method of detecting papillomavirus infection from a clinical sample of a human subject, comprising:
 obtaining a clinical sample from the human subject;   providing a papillomavirus detection composition of  claim 1 ;   conducting one or more immunological assays on the clinical sample; and   detecting the presence of one or more papillomavirus antibodies in the human subject using the recombinant papillomavirus protein.   
     
     
         8 . The method of  claim 7 , wherein the clinical sample is selected from the group consisting of cervical cells, cervical tissues, cervical swabs, body fluids, serum, blood, tumors, and combination thereof. 
     
     
         9 . The method of  claim 7 , wherein the one or more immunological assays are selected from the group consisting of ELISA assays, antigen assays for papillomavirus proteins, antibody assays for antibodies against papillomavirus proteins, assays for papillomavirus immunocomplexes, protein chip assays, radioimmunopercipitation assays, rapid membrane immunochromatographic assays, rapid stick immunochromatographic assays. 
     
     
         10 . The method of  claim 7 , wherein the recombinant papillomavirus protein is selected from the group consisting of recombinant HPV-16 E6 proteins, recombinant HPV-16 E7 proteins, recombinant HPV-18 E6 proteins, recombinant HPV-18 E7 proteins, recombinant HPV-16 L1 proteins, recombinant HPV-18 L1 proteins, and combinations thereof. 
     
     
         11 . A method of detecting papillomavirus infection from a clinical sample of a human subject, comprising:
 obtaining a clinical sample from the human subject;   providing a papillomavirus detection composition of  claim 1 ;   conducting one or more immunological assays on the clinical sample; and   detecting the presence of one or more papillomavirus viral proteins in the clinical sample of the human subject using anti-HPV antibody and the papillomavirus composition of  claim 1 .   
     
     
         12 . The method of  claim 11 , wherein the clinical sample is selected from the group consisting of cervical cells, cervical tissues, cervical swabs, body fluids, serum, blood, tumors, and combination thereof. 
     
     
         13 . The method of  claim 11 , wherein the one or more immunological assays are selected from the group consisting of ELISA assays, antigen assays for papillomavirus proteins, antibody assays for antibodies against papillomavirus proteins, assays for papillomavirus immunocomplexes, protein chip assays, radioimmunopercipitation assays, rapid membrane immunochromatographic assays, rapid stick immunochromatographic assays. 
     
     
         14 . The method of  claim 11 , wherein the recombinant papillomavirus protein is selected from the group consisting of recombinant HPV-16 E6 proteins, recombinant HPV-16 E7 proteins, recombinant HPV-18 E6 proteins, recombinant HPV-18 E7 proteins, recombinant HPV-16 L1 proteins, recombinant HPV-18 L1 proteins, and combinations thereof. 
     
     
         15 . Use of a papillomavirus detection composition of  claim 1 , for the detection of a disease consisting of HPV infection, cervical cancer, and combinations thereof.