Comparison of peptide enzyme-linked immunosorbent assay and radioimmunoprecipitation assay with in vitro-translated proteins for detection of serum antibodies to human papillomavirus type 16 E6 and E7 proteins.
Antibodies to human papilloma virus (HPV) type 16 (HPV-16) E6 and E7 proteins in serum are markers for HPV-associated invasive cervical carcinoma. We compared two assays, a radioimmunoprecipitation assay with in vitro-translated HPV-16 E6 and E7 proteins and an enzyme-linked immunosorbent assay (ELISA) with E6 and E7 synthetic peptides, for their abilities to discriminate serologically between patients with invasive cervical cancer and controls. Among the patients, antibody prevalences were higher by the E6 radioimmunoprecipitation assay (55.7%) than by the E6 peptide ELISA (15.5%), but among the controls, they were lower by the radioimmunoprecipitation assay (1.7%) than by the E6 peptide ELISA (5%). For E7, antibody prevalences among the patients were comparable by the radioimmunoprecipitation assay (43%) and the peptide ELISA (41%), but among the controls they were higher by the E7 peptide ELISA (17.4%) than by the radioimmunoprecipitation assay (4.1%). There was good agreement between the E7 radioimmunoprecipitation assay and the E7 peptide ELISA among patients but not among controls. In tests with representative sera, heat denaturation of the translated proteins resulted in a complete loss of reactivity to the E6 protein and a marked decrease in reactivity to the E7 protein. Our study showed that the radioimmunoprecipitation assay discriminates better than the peptide ELISA between patients with invasive cervical cancer and controls and that this is related to the ability of the radioimmunoprecipitation assay to detect conformational epitopes.
Sun
Y
Y
Department of Immunology and Infectious Diseases, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205.
Shah
K V
KV
Müller
M
M
Muñoz
N
N
Bosch
X F
XF
Viscidi
R P
RP
D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology*; D002277:Carcinoma / Q000473:pathology / Q000821:virology; D016022:Case-Control Studies; D003105:Colombia; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006358:Hot Temperature; D006801:Humans; D009361:Neoplasm Invasiveness; D009856:Oncogene Proteins, Viral / Q000276:immunology*; D027383:Papillomaviridae / Q000276:immunology* / Q000302:isolation & purification; D050725:Papillomavirus E7 Proteins; D030361:Papillomavirus Infections / Q000097:blood* / Q000276:immunology; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology*; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D011487:Protein Conformation; D011489:Protein Denaturation; D011863:Radioimmunoassay; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D012097:Repressor Proteins; D012680:Sensitivity and Specificity; D013030:Spain; D014412:Tumor Virus Infections / Q000097:blood* / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000473:pathology / Q000821:virology
D000914:Antibodies, Viral; D000956:Antigens, Viral; C068651:E6 protein, Human papillomavirus type 16; D000939:Epitopes; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16