Use of enzyme-labeled antigen for the detection of immunoglobulin M and A antibody to herpes simplex virus in serum and cerebrospinal fluid.

A direct enzyme-linked immunosorbent assay (ELISA that used peroxidase-labeled antigen) was developed for detection of IgM and IgA antibody to herpes simplex virus (HSV). The assay uses immuno-affinity-purified antihuman IgM or IgA antibody-coated wells of microtiter plates to separate IgM or IgA from other classes of antibody in serum or cerebrospinal fluid (CSF). The presence of specific IgM or IgA is detected by subsequent, consecutive incubation with peroxidase-labeled antigen and substrate. HSV antigen was purified by sucrose gradient centrifugation and coupled with peroxidase by the periodate method. By examining sucrose-gradient-fractionated sera the assays were shown to be specific for IgM and IgA classes of antibody. None of the sera from patients with Epstein-Barr virus (n = 20), cytomegalovirus (n = 20), or varicella-zoster virus (n = 8) infection or with both rheumatoid factor and IgG antibody to HSV (n = 13) reacted positively. Only one out of 78 sera from healthy persons was positive for IgA antibody to HSV, and none for IgM antibody. All 33 patients with HSV infection developed HSV-IgA, 22 developed HSV-IgM. Of the 11 patients with primary infection, all had IgM antibody in their first sera and six had IgA antibody. The corresponding figures for the 22 patients with recurrent infection were five and nine. Furthermore, HSV-IgA antibody was found in serum and CSF of all five patients with HSV encephalitis in the second week after onset of symptoms, indicating the usefulness of the assay as a noninvasive technique for diagnosing HSV encephalitis.