Four-antigen mixture containing v-cyclin for serological screening of human herpesvirus 8 infection.

Abstract

Improved diagnostic reagents and testing are currently needed for the serological detection of human herpesvirus 8 (HHV-8) infections. We evaluated the luciferase immunoprecipitation systems (LIPS) for profiling antibody responses to a panel of HHV-8 proteins for diagnosis of Kaposi sarcoma (KS)-infected individuals. Using a pilot serum set, LIPS detected robust antibody responses to several known antigens, and a screen of 14 additional HHV-8 proteins identified v-cyclin as a potentially new diagnostic antigen. In evaluating a training-serum set, a four-antigen panel (K8.1, v-cyclin, ORF65, and a LANA fragment) was found to provide sufficient information for diagnosis. Analysis of a validation serum set using the combined results from these four separate antigen tests showed 100% sensitivity and 100% specificity. Furthermore, a LIPS format using a mixture of the four antigens, which simplifies data collection and analysis, closely matched the diagnostic performance of the combined separate tests (R = 0.95). This four-antigen mixture format analyzed with the validation serum set also showed 100% sensitivity and 100% specificity but was not statistically different from two separate enzyme-linked immunosorbent assays (94% sensitivity and 100% specificity) using baculovirus-produced LANA and bacterially produced K8.1. Heat map analysis of KS patient antibody titers revealed marked heterogeneity in humoral responses to this four-antigen panel. Overall, the LIPS assay showed 97% sensitivity, and positive anti-v-cyclin antibodies were detected in approximately 75% of the KS sera. These results suggest that LIPS screening using an antigen mixture is a sensitive and high-throughput method for serological screening of HHV-8 infection in individuals with KS.

• Authors: Peter D Burbelo, Hannah P Leahy, Sandra Groot, Lisa R Bishop, Wendell Miley, Michael J Iadarola, Denise Whitby, Joseph A Kovacs
• Journal: Clinical and vaccine immunology : CVI (Clin Vaccine Immunol) Vol. 16 Issue 5 Pg. 621-7 (May 2009) ISSN: 1556-679X [Electronic] United States
• PMID: 19261774 (Publication Type: Evaluation Study, Journal Article, Research Support, N.I.H., Extramural, Research Support, N.I.H., Intramural)

Chemical References

• Antibodies, Viral
• Antigens, Viral

Topics

• Antibodies, Viral (blood)
• Antigens, Viral
• Herpesvirus 8, Human (immunology)
• Humans
• Mass Screening (methods)
• Sarcoma, Kaposi (diagnosis, virology)
• Sensitivity and Specificity