A sensitive enzyme-linked immunosorbent assay for the detection of herpes simplex virus antigens

J M Middeldorp, A M Hooymans, A J Kocken, A M van Loon, J A Emsbroek, R A Coutinho

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

A sandwich ELISA for the detection of herpes simplex virus (HSV) antigens was developed using sheep anti-HSV F(ab')2 fragments for capture and an indirect antibody system for detection. Current detection limits are 0.5 ng protein for HSV1 and 1.5 ng protein for HSV2. This compares to a single HSV1-infected Vero-cell in a background of 10(6) non-infected cells or 10 plaque forming units (PFU) of HSV1 in culture supernatants as determined in separate experiments. Limiting dilution experiments show that one PFU of HSV1 can be detected after overnight culture in both supernatant and cell extracts. The use of F(ab')2 for capture completely eliminated binding of Staphylococcus aureus. No cross-reactivity was observed with other human herpes viruses. When evaluated with 245 random 'left-overs' of genital swab specimens in transport medium the test showed a sensitivity and specificity of 77.2 and 97.8%, respectively, with respect to virus isolation in culture. In a preliminary study on 16 direct ELISA swab-specimens extracted in 0.5 ml ELISA sample buffer both sensitivity and specificity were 100% with respect to culture. In both clinical series there was a proportional relationship between the ELISA value and the estimated amount of infectious virus in the specimen.

Original languageEnglish
Pages (from-to)159-74
Number of pages16
JournalJournal of Virological Methods
Volume17
Issue number1-2
Publication statusPublished - Aug 1987

Cite this

Middeldorp, J. M., Hooymans, A. M., Kocken, A. J., van Loon, A. M., Emsbroek, J. A., & Coutinho, R. A. (1987). A sensitive enzyme-linked immunosorbent assay for the detection of herpes simplex virus antigens. Journal of Virological Methods, 17(1-2), 159-74.