AN ENZYME-LINKED IMMUNOSORBANT ASSAY FOR ANTIHERPES HERPES SIMPLEX VIRUS (HSV) AND ANTI-VARICELLA ZOSTER VIRUS (VZV) ANTIBODY (AB)

Abstract

The human newborn and immunocompromised host are particularly susceptible to the devastating effects of disseminated VZV and HSV infection. To further investigate the relevant immune parameters, we developed an ELISA for the detection of specific AB. Plastic microtiter plates (Dynatech)) were the solid phase for the antigen (Ag) (VZV/HSV antigen - Flow Labs), 100 ul/well of a 1:20 dilution in PBS. Following overnight (O/N) incubation Ag is replaced with PBS-gelatin (1 mg/ml) and incubated O/N 4°C. After washing thrice with PBS-tween 20 (.05%), the index sera or culture supernates are incubated 100 ul/well O/N 4C. Wells are washed as above and goat anti-human peroxidase conjugated Ab 1:2000 in PBS-gel-Tween-bovine -globulin (5 mg/ml) is incubated 2 hrs. at room temperature. After washing, substrate 100 ul/ml (ABTS-sigma, 1 mg/ml in phosphate buffer + .15% H₂O₂) is allowed to react at room temperature and read at 2 hrs. at 405 nm (Titertek Multiscan). We compared anti-VZV ELISA titers with those determined by FAMA technique:

The sensitivity of the method has been useful in the detection of Ab production by mononuclear cells in vitro.

Log in or create a free account to read this content

Gain free access to this article, as well as selected content from this journal and more on nature.com

Access through your institution

or

Sign in or create an account

Continue with Google

Continue with ORCiD