Four-antigen mixture containing v-cyclin for serological screening of human herpesvirus 8 infection.
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.
Burbelo
Peter D
PD
Neurobiology and Pain Therapeutics Section, Laboratory of Sensory Biology, National Institute of Dental and Craniofacial Research, Bethesda, Maryland 20892-44101, USA. burbelop@nidcr.nih.gov
Leahy
Hannah P
HP
Groot
Sandra
S
Bishop
Lisa R
LR
Miley
Wendell
W
Iadarola
Michael J
MJ
Whitby
Denise
D
Kovacs
Joseph A
JA
D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral; D019288:Herpesvirus 8, Human / Q000276:immunology*; D006801:Humans; D008403:Mass Screening / Q000379:methods*; D012514:Sarcoma, Kaposi / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D000956:Antigens, Viral