Development of molecular probes for simian herpesvirus detection

A number of serologic procedures are available for specifically determining past infection due to H. hominis, or H. simiae (B virus). A herpesvirus isolated from primate tissues, however, does not lend itself to specific identification. Procedures currently in routine use simply do not provide an unequivocal identification of the isolate. We have previously demonstrated that H. simiae contain polypeptide and glycoprotein antigens distinct from those of other herpesviruses. At selected intervals, infected cells and controls were harvested after being both pulsed as well as continuously labelled from 4-24 hours post infection with 3H-glucosamine. 3H-mannose, and 3H-L-amino acid mixture. Between 40-45 infected cell polypeptides were observed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) with 7-11 glycoproteins noted. Analysis of the bands by electroblot utilizing anti-B virus sera showed the presence of eleven reactive bands within this molecular weight range from preparations in infected Vero and HEp-2 cell lines. Glucosamine labelled major glycoproteins migrated at 130K, 100K, 95K, 65K, 39K; mannose moieties migrated at 95K and 65K. SDS-PAGE profiles of B-virus when reacted with anti-gA/gB from HSV-1 and HSV-2 showed weak reactivities on the 125-130K complex. Anti-VP5 from HSV-1 failed to indicate any reactivity. Three polypeptides were revealed by SDS-hydroxylapatite analysis of the 130K complex with approximate molecular weights of 115K, 123K, and 130K; the 115K and 123K bands reacted with antisera prepared against gA/gB from HSV-1 and HSV-2, but not to anti-IgC or anti-2gC.(ABSTRACT TRUNCATED AT 250 WORDS)