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Journal of Clinical Microbiology, February 2000, p. 696-701, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Use of Epidemiologically Well-Defined Subjects and Existing Immunofluorescence Assays To Calibrate a New Enzyme Immunoassay for Human Herpesvirus 8 Antibodies

Jeffrey N. Martin,1,2,* Zahwa Amad,3 Cynthia Cossen,3 Phung K. Lam,1 Dean H. Kedes,2,4,dagger Kimberly A. Page-Shafer,2 Dennis H. Osmond,1,2 and Bagher Forghani3

Department of Epidemiology and Biostatistics,1 and Department of Medicine,2 University of California, San Francisco, San Francisco, California 94105; Viral and Rickettsial Diseases Laboratory, California Department of Health Services, Berkeley, California 947043; and Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California 94143-04144

Received 13 August 1999/Returned for modification 24 September 1999/Accepted 12 November 1999

Agreement between assays for the detection of human herpesvirus 8 (HHV-8) antibodies has been limited. In part, this disagreement has been because assay calibration (i.e., differentiating positive from negative results) has not been done in a standardized fashion with reference to a wide spectrum of HHV-8-infected (true-positive) and HHV-8-uninfected (true-negative) persons. To describe the performance of an assay for HHV-8 antibodies more accurately, we used epidemiologically well-characterized subjects in conjunction with testing on two existing immunofluorescence assays for HHV-8 antibodies to define two groups: a group of 135 HHV-8-infected individuals (true positives), including Kaposi's sarcoma patients and those asymptomatically infected, and a group of 234 individuals with a high likelihood of being HHV-8 uninfected (true negatives). A new enzyme immunoassay (EIA), using lysed HHV-8 virion as the antigen target, was then developed. With the above true positives and true negatives as references, the sensitivity and specificity of the EIA associated with different cutoff values were determined. At the cutoff that maximized both sensitivity and specificity, sensitivity was 94% and specificity was 93%. When the EIA was used to test a separate validation group, a distribution of seropositivity that matched that predicted for the agent of Kaposi's sarcoma was observed: 55% of homosexual men were seropositive, versus 6% seropositivity in a group of children, women, and heterosexual men. It is proposed that the EIA has utility for large-scale use in a number of settings and that the calibration method described can be used for other assays, both to more accurately describe the performance of these assays and to permit more-valid interassay comparison.

* Corresponding author. Mailing address: Department of Epidemiology and Biostatistics, University of California, San Francisco, 74 New Montgomery, Suite 500, San Francisco, CA 94105. Phone: (415) 597-9219. Fax: (415) 597-9125. E-mail: martin{at}psg.ucsf.edu.

dagger Present address: Departments of Microbiology and Internal Medicine, Health Sciences Center, Charlottesville, VA 22908.

Journal of Clinical Microbiology, February 2000, p. 696-701, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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