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Comparative Study
. 1998 Jun;36(6):1574-7.
doi: 10.1128/JCM.36.6.1574-1577.1998.

Mapping and serodiagnostic application of a dominant epitope within the human herpesvirus 8 ORF 65-encoded protein

C P Pau  1 , L L Lam, T J Spira, J B Black, J A Stewart, P E Pellett, R A Respess
Affiliations
Comparative Study

Mapping and serodiagnostic application of a dominant epitope within the human herpesvirus 8 ORF 65-encoded protein

C P Pau et al. J Clin Microbiol. 1998 Jun.

Abstract

A dominant epitope within the human herpesvirus 8 (HHV8) ORF 65-encoded protein was mapped to an 8-amino-acid (aa) sequence (RKPPSGKK [aa 162 to 169]) by an amino acid replacement method. Using a 14-aa peptide (P4) encompassing this epitope as the antigen, we developed an enzyme immunoassay for HHV8 antibodies. The presence of P4 antibodies in a panel of 61 human serum specimens was highly correlated with biopsy-confirmed Kaposi's sarcoma. The homologous Epstein-Barr virus peptide derived from BFBR3-encoded protein did not interfere with the assay, suggesting that P4 is specific for HHV8.

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Figures

FIG. 1
FIG. 1
Seroreactivities of specimens from 20 KS+ HIV+ (I), 20 KS− HIV+ (II), and 20 KS− HIV− (III) persons with overlapping peptides P1, P2, P3, and P4 derived from the C-terminal half of the HHV8 ORF 65 gene product. The broken lines indicate the cutoff values, as defined by the mean baseline-corrected OD450 of the 20 KS− HIV− serum specimens plus 5 standard deviations.
FIG. 2
FIG. 2
Seroreactivities of three HHV8 antibody-positive serum specimens with P4 peptide analogs which differ from P4 by one residue. Substitutions and residue number corresponding to the HHV8 ORF 65 protein are shown in the x axis. A relative reactivity of 1.0 was assigned to peptide P4.
FIG. 3
FIG. 3
Seroreactivities of P4 with serum specimens (n = 6) preincubated with 0 to 10 μg/100 μl of competing peptides. P4 itself (a) and the P4 homolog derived from the C terminus of BFRF3-encoded EBV protein (b) were used as the competing peptide.
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