Direct immunofluorescence and direct immunoperoxidase staining were equally sensitive and specific for detection of herpes simplex virus antigen in lesion specimens, and each method showed 82% agreement with virus isolation results.

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Various subunit antigens of varicella-zoster (V-Z) virus were examined for complement-fixing (CF) activity with sera from homotypic infections and from herpes simplex virus (HSV) infections in which a CF antibody titer rise was demonstrated with crude V-Z antigen. The subunit antigens included nucleocapsids, envelopes, a soluble antigen produced from infected culture fluids by sucrose density gradient centrifugation, a soluble antigen produced by reducing the volume of clarified infected culture fluids, a soluble antigen derived from infected cell lysates, a "viral" antigen consisting largely of enveloped particles with a few nucleocapsids, and a cell membrane-associated antigen. None was more suitable than crude V-Z antigen for serodifferentiation of V-Z virus and HSV infections. The envelope antigen, cell membrane antigen, and the soluble antigen prepared by density gradient centrifugation showed little reactivity with sera from varicella and HSV infections, but gave high antibody titers with sera from zoster infections, suggesting that a secondary V-Z virus infection is required to produce an antibody response to these subunit antigens. Patients with varicella and zoster infections and the selected patients with HSV infections all showed significant CF antibody responses to the other V-Z subunit antigens.It is well established that infection with herpes simplex virus (HSV) may elicit heterotypic antibody responses to varicella-zoster (V-Z) virus in individuals who have experienced a previous V-Z virus infection (3,8,9,11). Differential serological diagnosis of HSV and V-Z virus infections may be hampered by these heterotypic antibody responses, which are demonstrable by fluorescent-antibody staining and neutralization tests (8,9) as well as by complement fixation (CF) tests (3,9,11).Martin and Palmer (5) reported that a cellfree soluble CF antigen derived from V-Z virusinfected cell cultures failed to show heterotypic reactivity with sera from HSV patients who showed CF antibody titer rises with crude V-Z antigens, and suggested that such soluble V-Z CF antigens might be useful in the differential serological diagnosis of HSV and V-Z virus infections. The present studies were undertaken to explore further the CF reactivity of subunit antigens of V-Z virus with sera from homotypic infections and from HSV infections, and to assess the usefulness of soluble V-Z antigens for serological differentiation of HSV and V-Z virus infections. MATERIALS AND METHODSPreparations of CF antigens. The Batson strain of V-Z virus (10) and the L 645 strain of human fetal diploid lung (HFDL) cells, established by J. H. Schieble of this laboratory, were used for preparation of V-Z virus antigens. Infected HFDL cells in a 29-oz (ca. 800-ml) culture bottle which showed a 2+ viral cytopathic effect were dispersed with trypsin and used to infect six roller bottle cultures of HFDL cells; these were maintained on Eagle minimal essential medium supplemented with 2% inactivated fetal bovine serum. Cultures were harvested after 4 days of inc...

A hemagglutination-inhibition (HI) test for rubella is described which utilizes human group 0, rather than 1-day-old chick, erythrocytes. The test was found to be as sensitive and reproducible for detection of rubella antibody as HI tests employing chick erythrocytes. Advantages to the use of human erythrocytes are (i) they are more available, (ii) it is unnecessary to absorb natural agglutinins from human test sera, and (iii) heparin-MnCl2-treated sera do not agglutinate human erythrocytes, as is sometimes the case with chick erythrocytes. Factors influencing the reliability of the test are discussed.