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Clinical and Diagnostic Laboratory Immunology, May 2002, p. 577-582, Vol. 9, No. 3
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.3.577-582.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Optimization of a Human Papillomavirus-Specific Enzyme-Linked Immunosorbent Assay

Kevin L. Karem,* Alysia C. Poon, Cynthia Bierl, Rosane Nisenbaum, and Elizabeth Unger

Viral Exanthems and Herpesvirus Branch, Division of Viral and Rickettsial Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333

Received 20 December 2001/ Returned for modification 31 January 2002/ Accepted 21 February 2002

A strategy was developed for the control, standardization, and critical evaluation of an enzyme-linked immunosorbent assay (ELISA) for the detection of human papillomavirus-specific immunoglobulin G in human sera. Control human sera, polyclonal animal sera, and monoclonal antibodies were used to establish optimal assay parameters, including antigen coating, serum dilutions, and criteria for daily reproducibility, monitoring, and rejection of assays. Three evaluation techniques were used in parallel to define an optimal cutoff absorbance value that yields greater than 93% sensitivity and 98.5% specificity in the assay's ability to discriminate positive and negative control sera. This strategy provides an optimal method by which to determine cutoff absorbance values for ELISA.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, Division of Viral and Rickettsial Disease, Viral Exanthems and Herpesvirus Branch, 1600 Clifton Rd., Mail Stop G-41, Atlanta, GA 30333. Phone: (404) 639-2179. Fax: (404) 639-3540. E-mail: kkarem{at}cdc.gov.

Clinical and Diagnostic Laboratory Immunology, May 2002, p. 577-582, Vol. 9, No. 3
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.3.577-582.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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