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Journal of Clinical Microbiology, May 2002, p. 1755-1760, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1755-1760.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Enhanced Enzyme-Linked Immunosorbent Assay for Detection of Antibodies to Virus-Like Particles of Human Papillomavirus

Yevgeniy Y. Studentsov,1 Mark Schiffman,2 Howard D. Strickler,3 Gloria Y. F. Ho,3 Yuk-Ying Susana Pang,4 John Schiller,4 Rolando Herrero,5 and Robert D. Burk1,3,6*

Department of Microbiology & Immunology,1 Department of Epidemiology & Social Medicine,3 Departments of Pediatrics and Obstetrics & Gynecology, Albert Einstein College of Medicine, Bronx, New York,6 Division of Cancer Epidemiology and Genetics,2 Laboratory of Cellular Oncology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland,4 Proyecto Epidemiologico Guanacaste, San Jose, Costa Rica5

Received 30 November 2001/ Returned for modification 9 February 2002/ Accepted 20 February 2002

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.

* Corresponding author. Mailing address: Cancer Research Center, Albert Einstein College of Medicine, Ullmann Bldg., Rm. 515, Bronx, NY 10461. Phone: (718) 430-3744. Fax: (718) 430-8975. E-mail: burk{at}aecom.yu.edu.

Journal of Clinical Microbiology, May 2002, p. 1755-1760, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1755-1760.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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