Heparin-based ELISA reduces background reactivity in virus-like particle-based papillomavirus serology.
The interaction between human papillomavirus (HPV) particles and cell surface heparan sulfate requires intact conformation of the HPV particles. Type-specific HPV serology is currently based on virus-like particles (VLPs) with intact conformation. Presence of incorrectly folded VLPs in VLP preparations is recognized as an important cause of cross-reactivity in HPV serology. Heparin-coated microtitre plates were evaluated for capturing conformationally correct VLPs and improving the type specificity of HPV serology. Hybrid VLPs between HPV16 and HPV11, which had been found to have significant reactivity with children's sera and a batch of HPV18 VLPs that had failed the quality control because of significant reactivity with sera from virginal women, were tested in parallel with heparin ELISA, ordinary ELISA and type-specific mAb capture ELISA. Control sera from children that had detectable reactivity with HPV16/11 hybrid VLPs in ordinary ELISA did not react in heparin-based ELISA, but some hybrid VLPs also had background reactivity in capture ELISAs. Control sera from virginal women that had some reactivity with a poor quality HPV18 VLP preparation in ordinary ELISA had no reactivity in heparin or capture ELISA, suggesting that certain VLP preparations expose cross-reactive epitopes that are not exposed on VLPs with heparin-binding ability. As the sensitivity was similar or only marginally affected by the use of heparin plates, use of heparin-coated plates may improve the type specificity of VLP-based ELISAs and reduce interassay variability attributable to variable quality of different VLP batches.
Wang
Xiaohong
X
Department of Medical Microbiology, University Hospital, 20502 Malmö, Sweden.
Sapp
Martin
M
Institute of Medical Microbiology and Hygiene, University of Mainz, 55101 Mainz, Germany.
Christensen
Neil D
ND
Department of Microbiology and Immunology, The Milton S. Hershey Center, 500 University Drive, Hershey, PA 17033, USA.
Dillner
Joakim
J
Department of Medical Microbiology, University Hospital, 20502 Malmö, Sweden.
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006493:Heparin; D006801:Humans; D008297:Male; D008875:Middle Aged; D027383:Papillomaviridae / Q000276:immunology*; D030361:Papillomavirus Infections / Q000097:blood / Q000175:diagnosis*; D011786:Quality Control; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D006493:Heparin