Enhanced enzyme-linked immunosorbent assay for detection of antibodies to virus-like particles of human papillomavirus.
Measurement of antibodies to human papillomavirus (HPV) is complicated by many factors. Although enzyme-linked immunosorbent assays (ELISAs) that use virus-like particles (VLPs) have proved useful, the assays have, in general, had moderate sensitivities and low signal-to-noise ratios. To enhance the performance of the assay, a systematic investigation was undertaken to examine key variables used in ELISAs for the detection of antibodies to VLPs of HPV. Incorporation of two vinyl polymers, polyvinyl alcohol (molecular weight, 50,000) (PVA-50) and polyvinylpyrrolidone (molecular weight, 360,000) (PVP-360), was found to increase the sensitivity as well as the specificity of the assay for the detection of antibodies to VLPs of HPV. In particular, the addition of PVA-50 to the blocking solution reduced the amount of nonspecific binding of antibodies to VLPs and the microplate surface, whereas the addition of PVP-360 increased the sensitivity of antibody detection. The new ELISA demonstrated increased sensitivity and specificity for the detection of cervical HPV type 16 infection compared to those of a prototype assay with coded clinical serum samples from women with known cervicovaginal HPV infection status. It is anticipated that the enhanced ELISA conditions will have wide application to a large number of clinical diagnostic assays.
Studentsov
Yevgeniy Y
YY
Department of Microbiology & Immunology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Schiffman
Mark
M
Strickler
Howard D
HD
Ho
Gloria Y F
GY
Pang
Yuk-Ying Susana
YY
Schiller
John
J
Herrero
Rolando
R
Burk
Robert D
RD
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D004279:DNA, Viral / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007202:Indicators and Reagents; D008892:Milk / Q000821:virology; D027383:Papillomaviridae / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D015203:Reproducibility of Results
D000914:Antibodies, Viral; D004279:DNA, Viral; D007202:Indicators and Reagents