Simultaneous detection of and differentiation between herpes simplex and varicella-zoster viruses with two fluorescent probes in the same test system

Abstract

Specimens from skin lesions were examined simultaneously for herpes simplex virus (HSV) and varicellazoster virus (VZV) by direct specimen testing and shell vial culture in single-test systems. For direct testing, cells in a single specimen well were stained with a combination direct-indirect immunofluorescence stain by using two fluorescent tags. A total of 203 fresh specimens were tested in parallel. Of these, 100 specimens contained too few cells for the direct VZV comparison and 91 contained too few cells for the HSV comparison. After these specimens were eliminated, the sensitivities and specificities, respectively, of the dual direct test were 86.1 and 97.3% for HSV compared with single culture and 92.2 and 100% for VZV compared with single direct testing. Shell vial monolayers in the combined cultures were stained for both viruses by the same method. A total of 305 fresh specimens were cultured in parallel by dual-and single-culture methods. The sensitivities and specificities, respectively, of the combined culture compared with separate cultures were 100 and 98.4% for HSV and 87.9 and 99.2% for VZV. The combined methods gave a performance comparable to those of single tests, required less specimen volume, and were less costly to perform.

Laboratory diagnosis of viral infection

“…If the screening reagent is positive, additional slides are stained to identify the specific pathogen, using either single fluorescein-labeled antibodies or dualantibody reagents with two different fluorophore labels. Dual fluorophore-labeled antibody pools can also be used to detect and differentiate between HSV and VZV (Brumback et al, 1993).…”

Differential diagnoses of influenza A virus, influenza B virus, and respiratory syncytial virus infections by direct immunofluorescence using mixtures of monoclonal antibodies of different isotypes

“…It would be desirable to evaluate the same cells for different viral antigens. This problem has been addressed for other viruses by using a combination of direct and indirect fluorescent-antibody procedures and by using different fluorescent probes (1,2). Our approach was to exploit the differences in isotypes between monoclonal antibodies (MAbs) that are used for the diagnosis of influenza A and B viruses and RSV in a rapid DIF method (11).…”